anticd74 (R&D Systems)

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R&D Systems anticd74

Fig. 5. IRF8 control of components of the MHCII complex. (A) FACS analysis of H2-IA/IE (left) and HLA-DR (right) in models of IRF8 KO; WB of IRF8 in RIVA and SU-DHL2 KO models, and WB of MHCII in all IRF8 KO models in DLBCL. (B) FACS of <t>CD74</t> in models of IRF8 KO. (C) FACS of H2-DM and HLA-DM in IRF8 KO models. (D) Left: FACS of CD74 and H2-DM in the IRF8 KO A20 lymphoma model “rescued” with IRF8 WT or missense or nonsense mutants (top and bottom). Right: FACS of CD74 and H2-DM in the IRF8 KO 2PK-3 lymphoma model “rescued” with IRF8 WT or missense and nonsense mutants (top and bottom). (E) Top: ChIP-qPCR of IRF8 binding to the indicated promot- ers – controls are IgG pull down, and a genomic region without a predicted IRF8 binding site (neg ctrl). Bottom: ChIP-qPCR of IRF8 WT, N87Y, or I424T binding to the Cd74, H2-Dm, Ciita, or H2-Aa promoters. (F) Top: WB of CD74 in 2PK-3 and A20 CD74-KO models. Bottom: IL-2 levels and % of CD4/CD25+ cells in IRF8/CD74 WT, IRF8 KO, or CD74 KOs models. (G) Left to right: A20, 2PK-3, and BCL1 models of IRF8 KO with CD74 ectopic expression (ee). WB of CD74-FLAG, IL-2 levels and % of CD4/CD25+ cells in IRF8/ CD74 WT, IRF8 KO, or IRF8KO + CD74. (H) Left: WB of CD74-FLAG in IRF8 WT, N87Y, and I424T A20 models. Right: IL-2 levels and % of CD4/CD25+ cells in IRF8 WT, N87Y, and I424T (−/+ CD74 ectopic expression) models. Data are means ± SD of three biological replicates. FACS displayed as relative mean fluorescence intensity (MFI). P values are from ANOVA, with Bonferroni or Fisher’s LSD posttest, or two-sided Student’s t test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Anticd74, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anticd74/Mouse+CD74+Antibody/pm38996030-370-39-49
Average 93 stars, based on 18 article reviews

anticd74 - by Bioz Stars, 2026-10

93/100 stars

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1) Product Images from "IRF8-mutant B cell lymphoma evades immunity through a CD74-dependent deregulation of antigen processing and presentation in MHCII complexes."

Article Title: IRF8-mutant B cell lymphoma evades immunity through a CD74-dependent deregulation of antigen processing and presentation in MHCII complexes.

Journal: Science advances

doi: 10.1126/sciadv.adk2091

Figure Legend Snippet: Fig. 5. IRF8 control of components of the MHCII complex. (A) FACS analysis of H2-IA/IE (left) and HLA-DR (right) in models of IRF8 KO; WB of IRF8 in RIVA and SU-DHL2 KO models, and WB of MHCII in all IRF8 KO models in DLBCL. (B) FACS of CD74 in models of IRF8 KO. (C) FACS of H2-DM and HLA-DM in IRF8 KO models. (D) Left: FACS of CD74 and H2-DM in the IRF8 KO A20 lymphoma model “rescued” with IRF8 WT or missense or nonsense mutants (top and bottom). Right: FACS of CD74 and H2-DM in the IRF8 KO 2PK-3 lymphoma model “rescued” with IRF8 WT or missense and nonsense mutants (top and bottom). (E) Top: ChIP-qPCR of IRF8 binding to the indicated promot- ers – controls are IgG pull down, and a genomic region without a predicted IRF8 binding site (neg ctrl). Bottom: ChIP-qPCR of IRF8 WT, N87Y, or I424T binding to the Cd74, H2-Dm, Ciita, or H2-Aa promoters. (F) Top: WB of CD74 in 2PK-3 and A20 CD74-KO models. Bottom: IL-2 levels and % of CD4/CD25+ cells in IRF8/CD74 WT, IRF8 KO, or CD74 KOs models. (G) Left to right: A20, 2PK-3, and BCL1 models of IRF8 KO with CD74 ectopic expression (ee). WB of CD74-FLAG, IL-2 levels and % of CD4/CD25+ cells in IRF8/ CD74 WT, IRF8 KO, or IRF8KO + CD74. (H) Left: WB of CD74-FLAG in IRF8 WT, N87Y, and I424T A20 models. Right: IL-2 levels and % of CD4/CD25+ cells in IRF8 WT, N87Y, and I424T (−/+ CD74 ectopic expression) models. Data are means ± SD of three biological replicates. FACS displayed as relative mean fluorescence intensity (MFI). P values are from ANOVA, with Bonferroni or Fisher’s LSD posttest, or two-sided Student’s t test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Techniques Used: Control, ChIP-qPCR, Binding Assay, Expressing, Fluorescence

Fig. 6. IRF8 effects on B cell lymphoma aggressiveness and immune microenvironment. (A) Growth curve of lymphomas expressing IRF8 WT, N87Y, Q392X, or I424T. (B) FACS-based quantification of CD3, CD4 and CD8 T cells in the TME of IRF8 WT or mutant lymphomas. (C) FACS-based quantification of Tregs and NK cells in the TME of IRF8 WT or mutant lymphomas. (D) IHC-based quantification of T cell infiltrate in B cell lymphomas expressing IRF8 WT, N87Y, or I424T. Representative staining (B220, pink; CD3, brown) is shown to the right, scale bar is displayed. (E) Growth curve of lymphomas expressing IRF8 WT, IRF8 N87Y or IRF8 I424T (−/+ CD74 expression). (F) FACS- based quantification of CD3, CD4 and CD8 in the TME of IRF8 WT or mutant lymphomas (−/+ CD74 expression). (G) FACS-based quantification of Tregs and NK cells in the TME of IRF8 WT or mutant lymphomas (−/+ CD74 expression). (H) TH1/TH2 ratio, TH1, TH2, and TFH cells in the TME of IRF8 WT or mutant lymphomas (−/+ CD74 expression). (I) TH1/TH2 ratio and TFH cells in the TME of IRF8 WT, missense (N87Y) or truncating (Q392X) mutant lymphomas. (J) Growth curve of lymphomas models expressing IRF8 WT or N87Y in mice treated with control antibody or anti–PD-L1 antibody; FACS-based quantification of CD4 and CD8 in IRF8 N87Y lymphomas treated with control or anti–PD-L1 antibody. For all panels, data are means ± SD of multiple independent cohorts (n indicated in the figure). P values are from one-way ANOVA with Fisher’s LSD posttest, Mann-Whitney test, or two-sided Student’s t test; *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001.

Figure Legend Snippet: Fig. 6. IRF8 effects on B cell lymphoma aggressiveness and immune microenvironment. (A) Growth curve of lymphomas expressing IRF8 WT, N87Y, Q392X, or I424T. (B) FACS-based quantification of CD3, CD4 and CD8 T cells in the TME of IRF8 WT or mutant lymphomas. (C) FACS-based quantification of Tregs and NK cells in the TME of IRF8 WT or mutant lymphomas. (D) IHC-based quantification of T cell infiltrate in B cell lymphomas expressing IRF8 WT, N87Y, or I424T. Representative staining (B220, pink; CD3, brown) is shown to the right, scale bar is displayed. (E) Growth curve of lymphomas expressing IRF8 WT, IRF8 N87Y or IRF8 I424T (−/+ CD74 expression). (F) FACS- based quantification of CD3, CD4 and CD8 in the TME of IRF8 WT or mutant lymphomas (−/+ CD74 expression). (G) FACS-based quantification of Tregs and NK cells in the TME of IRF8 WT or mutant lymphomas (−/+ CD74 expression). (H) TH1/TH2 ratio, TH1, TH2, and TFH cells in the TME of IRF8 WT or mutant lymphomas (−/+ CD74 expression). (I) TH1/TH2 ratio and TFH cells in the TME of IRF8 WT, missense (N87Y) or truncating (Q392X) mutant lymphomas. (J) Growth curve of lymphomas models expressing IRF8 WT or N87Y in mice treated with control antibody or anti–PD-L1 antibody; FACS-based quantification of CD4 and CD8 in IRF8 N87Y lymphomas treated with control or anti–PD-L1 antibody. For all panels, data are means ± SD of multiple independent cohorts (n indicated in the figure). P values are from one-way ANOVA with Fisher’s LSD posttest, Mann-Whitney test, or two-sided Student’s t test; *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001.

Techniques Used: Expressing, Mutagenesis, Staining, Control, MANN-WHITNEY

Control:

Article Title: IRF8-mutant B cell lymphoma evades immunity through a CD74-dependent deregulation of antigen processing and presentation in MHCII complexes.
Article Snippet: Before electrophoresis, samples were denatured in 5× laemmli buffer (8% SDS, 0.4 M DTT, 0.2 M tris- HCl, 4.3 M glycerol, and 6 mM bromophenol blue) and separated by SDS–polyacrylamide gel electrophoresis (SDS- PAGE), transferred to immobilon- P polyvinylidene difluoride (PVDF) membrane (Millipore) at 100 V for 1 to 2 hours at 4°C.(PVDF) membrane (Millipore) at 100 V for 1 to 2 hours at 4°C. ... Membranes were blocked for 1 hour and probed with 5% nonfat dry milk with the following primary antibodies: anti- IRF8 [clone D20D8, catalog no. 5628, Cell Signaling Technology and clone E- 9, catalog no. sc- 365042, Santa Cruz Biotechnology)], antiCD74 (sheep anti- mouse cd74, polyclonal antibody, catalog no. AF7478, R&D Systems), anti- PU1 (clone 9G7, catalog no. 2258, Cell Signaling Technology), anti- FLAG (clone 9A3, catalog no. 8146, Cell Signaling Technology), anti–MHC class II (clone LGII- 612.14, catalog no. 68258, Cell Signaling Technology), anti- CIITA antibodies (rabbit polyclonal, catalog no. 3793, Cell Signaling Technology, and mouse monoclonal, clone 7- 1H, sc- 13556, Santa Cruz Biotechnology), and anti–β- actin (#A2228, Sigma- Aldrich).. Membranes were developed with the SuperSignal West Pico PLUS Chemiluminescent Substrate (Thermo Fisher Scientific) and captured on x- ray film.Membranes were developed with the SuperSignal West Pico PLUS Chemiluminescent Substrate (Thermo Fisher..

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anticd74 (R&D Systems)

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