# cd74 (Becton Dickinson)

## Structured Review

Cd74, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more

https://www.bioz.com/result/cd74/product/Becton Dickinson

Average 99 stars, based on 1 article reviews

Price from $9.99 to $1999.99

## Images

### 1) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 2) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 3) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 4) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 5) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 6) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 7) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 8) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 9) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 10) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 11) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 12) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 13) Product Images from "17?-Estradiol Inhibits Wound Healing in Male Mice via Estrogen Receptor-?"

**Article Title: **17?-Estradiol Inhibits Wound Healing in Male Mice via Estrogen Receptor-?

**Journal: **The American Journal of Pathology

**doi: **10.2353/ajpath.2010.090432

**Figure Legend Snippet:**Effects of chronic 17β-estradiol on wound inflammation in castrated mice. A: Immunostaining for the neutrophil marker Ly6G, and overall neutrophil numbers, in day three wounds from castrated (CSX, CX) MF1 mice, some treated with 17β-estradiol (E2) from days −14 to +3 (CSX +E2, CXE). B: Immunostaining for the macrophage marker Mac3, and numbers of macrophages, in day three wounds from E2-treated CSX MF1 mice. C: Immunostaining for arginase 1, a marker of AA macrophages, and day three wound numbers of arginase 1-positive cells, in E2-treated CSX MF1 mice. Immunostaining for TNF-α ( D ), MIF ( E ), and CD74 ( F ) in day three wounds from E2-treated CSX mice. G: Day three wound expression of the Tnfa , Mif , and Cd74 genes, determined by qPCR. H: Cellular levels of CD74 protein in peritoneal macrophages activated with LPS (+) for 2 hours (Con) and then treated with 100 nmol/L E2 or left untreated (Con) for a further three hours (immunoblotting). I: Expression of the Cd74 gene in control and 100 nmol/L E2-treated macrophages (qPCR). Statistical significance ( C and F ) was determined by unpaired Student’s t -tests: * P

**Techniques Used: **Mouse Assay, Immunostaining, Marker, Expressing, Real-time Polymerase Chain Reaction

### 14) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 15) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 16) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 17) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 18) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 19) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 20) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 21) Product Images from "Lipidated promiscuous peptide augments the expression of MHC-II molecules on dendritic cells and activates T cells"

**Article Title: **Lipidated promiscuous peptide augments the expression of MHC-II molecules on dendritic cells and activates T cells

**Journal: **The Indian Journal of Medical Research

**doi: **

**Figure Legend Snippet:**L91 induces MHC-II but downregulates CD74 expression. Bone marrow derived cells (BMDCs) were cultured in medium containing L91, F91 or lipopolysaccharides (LPS). The CD11c + cells were analyzed by flowcytometry for the expression of (a) MHC-II; (b) CD74 (immature MHC-II). Numbers in the flow cytometry histograms indicate mean fluorescence intensity (MFI) and are representative of 3 independent experiments; (c) bar graphs depict the change in the ratio of MHC-II to CD74. The cells cultured with medium alone and with LPS were used as negative and positive controls, respectively. Data are shown as the mean±SD are from 3 independent experiments. *** P

**Techniques Used: **Expressing, Derivative Assay, Cell Culture, Flow Cytometry, Cytometry, Fluorescence

### 22) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 23) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 24) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 25) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 26) Product Images from "The asparaginyl endopeptidase legumain after experimental stroke"

**Article Title: **The asparaginyl endopeptidase legumain after experimental stroke

**Journal: **Journal of Cerebral Blood Flow & Metabolism

**doi: **10.1038/jcbfm.2010.39

**Figure Legend Snippet:**Analysis of CD74 positive cells in the ischemic hemisphere of legumain (lgmn) wild-type and deficient mice after transient middle cerebral artery occlusion (tMCAO). Quantification of CD74 + cells from the ischemic hemisphere of lgmn wild-type (

**Techniques Used: **Mouse Assay

### 27) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 28) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 29) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 30) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 31) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 32) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 33) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 34) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 35) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 36) Product Images from "Milatuzumab-Conjugated Liposomes as Targeted Dexamethasone Carriers for Therapeutic Delivery in CD74+ B-cell Malignancies"

**Article Title: **Milatuzumab-Conjugated Liposomes as Targeted Dexamethasone Carriers for Therapeutic Delivery in CD74+ B-cell Malignancies

**Journal: **Clinical cancer research : an official journal of the American Association for Cancer Research

**doi: **10.1158/1078-0432.CCR-12-2046

**Figure Legend Snippet:**Localization of CD74-ILs in target cells. CD74-ILs, after 1-hour incubation with Raji cells (A, B, G, J) visualized by confocal microscopy. CD74-ILs are observed inside the cells and also on the cell membrane. Controls such as IgG-ILs (C and L) and nontargeted liposomes (D, E, K) did not enter/bind the cells. CD74-ILs in CD74 − cell line Jurkat (I) did not bind/enter the cells. All lipids are R18-labeled.

**Techniques Used: **Incubation, Confocal Microscopy, Labeling

**Figure Legend Snippet:**CD74-IL-DEX decreases viability and mitochondrial activity in CLL cells. Primary CLL cells ( n = 14) were treated for 24 hours with dexamethasone (10 μmol/L) in free or liposomal forms with cross-linked milatuzumab (CD74+Fc) and all CD74 concentrations at 5 μg/mL (A). Flow cytometric analysis of percentage of PI-positive cells indicated that CD74-IL-DEX kills significantly more B-CLL cells than CD74-ILs ( P

**Techniques Used: **Activity Assay, Flow Cytometry

**Figure Legend Snippet:**CD74-IL-DEX and L-DEX increase cellular GilZ level. Immunoblot analysis of GilZ induction in whole-cell lysates isolated from Raji cells treated with different formulations of DEX at 10 μmol/L and equivalent CD74.

**Techniques Used: **Isolation

**Figure Legend Snippet:**CD74-ILs bind to and are internalized into CD74 + Raji cells. CD74-ILs labeled with calcein are shown by flow cytometry to bind to CD74( + ) Raji B cells (A) but not CD74 − Jurkat T cells (B). Nonspecific IgG-ILs do not bind to Raji cells (AandB). Internalization of CD74-ILs and anti-CD74 ( n = 3) is shown in Raji cells (C), over time portrayed by change in MFI. The results were normalized to IgG isotype and IgG-ILs.

**Techniques Used: **Labeling, Flow Cytometry, Cytometry

### 37) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 38) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 39) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

### 40) Product Images from "CD74 is a regulator of hematopoietic stem cell maintenance"

**Article Title: **CD74 is a regulator of hematopoietic stem cell maintenance

**Journal: **PLoS Biology

**doi: **10.1371/journal.pbio.3001121

**Figure Legend Snippet:**CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

**Figure Legend Snippet:**Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

**Figure Legend Snippet:**CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

**Techniques Used: **Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

**Figure Legend Snippet:**Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

**Techniques Used: **FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

**Techniques Used: **Irradiation, Mouse Assay, Two Tailed Test

**Figure Legend Snippet:**CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

**Techniques Used: **Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

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