Journal: The Journal of Experimental Medicine

Article Title: Dietary restriction improves repopulation but impairs lymphoid differentiation capacity of hematopoietic stem cells in early aging

doi: 10.1084/jem.20151100

Figure Lengend Snippet: IGF1 injection reverts the increases in HSC quiescence but not the suppression of lymphopoiesis in mice exposed to DR. (A) ELISA-determined serum level of IGF1 in mice treated with DR or AL for 4 d or refed with an AL diet for 3 d after 4 d of DR ( n = 3–4 mice per group; n = 2 independent experiments). (B) ELISA-determined serum level of IGF1 in 2-mo-old p53 −/− mice and p53 +/+ mice (littermates) that were treated with DR or AL for 2 wk ( n = 3–4 mice per group; n = 2 independent experiments). (C–G) Mice were treated with a DR or AL diet for 8 d. In parallel, recombinant human IGF1 protein or vehicle control was subcutaneously injected twice per day at a dose of 500 µg/kg ( n = 4 mice per group; n = 2 independent experiments). FACS analysis was performed 24 h after the last injection on freshly isolated BM cells: percentage of HSCs in the indicated cell cycle stages (C), total numbers of CLPs (D), pro–B cells (E), MEPs (F), and proEry’s (G) per million total BM cells are show. AL con, control-injected AL mice; AL IGF1, IGF1-injected AL mice; DR con, control-injected DR mice; DR IGF1, IGF1-injected DR mice. Data are displayed as mean ± SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 by one-way ANOVA. ns, not significant.

Article Snippet: ELISA for detecting IGF1, IL-6, IL-7, and total corticosterone was performed using the IGF-I mouse/rat ELISA kit (Mediagnost), the Mouse IL-6 Quantikine ELISA kit (R&D Systems), the Mouse IL-7 Quantikine ELISA kit (R&D Systems), and Corticosterone EIA kit (ARBOR ASSAYS), respectively, according to the manufacturer’s instructions.

Techniques: Injection, Enzyme-linked Immunosorbent Assay, Recombinant, Control, Isolation

Journal: The Journal of Experimental Medicine

Article Title: Dietary restriction improves repopulation but impairs lymphoid differentiation capacity of hematopoietic stem cells in early aging

doi: 10.1084/jem.20151100

Figure Lengend Snippet: IL-7 or IL-6 injection reverts suppression of lymphopoiesis in mice exposed to DR. (A) ELISA-determined serum level of IL-7 in mice treated with DR or AL for 7 d or refed with AL diet for 3 d after 7 d of DR ( n = 3–4 mice per group; n = 2 independent experiments). One-way ANOVA. (B–G) 2-mo-old mice were treated with DR or AL diet for 9 d. In parallel, mouse IL-7 protein or vehicle control was subcutaneously injected for 9 d at a dose of 50 µg/kg once per day ( n = 5 mice per group; n = 2 independent experiments). Mice were sacrificed for analysis 24 h after the last injection. (B–D) FACS analysis was performed on freshly isolated BM cells to determine the total number of CLPs (B) and MEPs (C), and the percentage of CLPs (D) of DR mice in the indicated phases of the cell cycle (FACS analysis by Ki67 and DAPI staining). (E and F) the ratio of CLPs versus lymphoid-biased HSCs (E) and of pro–B cells versus CLPs (F). (G) The mRNA expression levels of genes that are associated with lymphoid cell differentiation and erythroid cell differentiation were determined by qRT-PCR in freshly isolated lymphoid-biased HSCs under the indicated treatment conditions. One-way ANOVA (B, C, and E–G) or unpaired two-tailed Student’s t test (D) was used. (H) ELISA-determined serum level of IL-6 in mice treated with DR or AL for 7 d or refed with AL diet for 3 d after 7 d of DR ( n = 3–4 mice per group; n = 2 independent experiments). One-way ANOVA was used. (I) 2-mo-old mice were treated with a DR or AL diet for 5 d. In parallel, mouse IL-6 protein or vehicle control was subcutaneously injected into mice daily at a dose of 50 µg/kg ( n = 4 mice per group; n = 2 independent experiments). The total number of CLPs was determined by FACS analysis, 24 h after the last injection. One-way ANOVA analysis. (J–P) ADX or sham surgery was performed on 2-mo-old mice. 10 d after operation, mice were treated with DR or AL for 9 d ( n = 4–5 mice per group; n = 2 independent experiments). Mice of the indicated cohorts were analyzed as follows: (J, O, and P) ELISA-determined serum levels of total corticosterone (J), IL-7 (O), and IL-6 (P); (K–M) FACS analysis of the total number of CLPs (K) and MEPs (M) per million BM cells and of the percentage of CLPs (L) of DR mice in different stages of the cell cycle (by Ki67 and DAPI staining); (N) expression of lymphoid- and erythroid-specific genes in lymphoid-biased HSCs of mice of the indicated cohorts. One-way ANOVA (J, K, and M–P) or unpaired two-tailed Student’s t test (L) was used. AL con, control-injected AL mice or sham-operated AL mice; AL IL-7, IL-7–injected AL mice; DR con, control-injected DR mice or sham-operated DR mice; DR IL-7, IL-7–injected DR mice; AL IL-6, IL-6–injected AL mice; DR IL-6, IL-6–injected DR mice; AL ADX, AL mice with ADX; DR ADX, DR mice with ADX. Data are displayed as mean ± SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. ns, not significant.

Article Snippet: ELISA for detecting IGF1, IL-6, IL-7, and total corticosterone was performed using the IGF-I mouse/rat ELISA kit (Mediagnost), the Mouse IL-6 Quantikine ELISA kit (R&D Systems), the Mouse IL-7 Quantikine ELISA kit (R&D Systems), and Corticosterone EIA kit (ARBOR ASSAYS), respectively, according to the manufacturer’s instructions.

Techniques: Injection, Enzyme-linked Immunosorbent Assay, Control, Isolation, Staining, Expressing, Cell Differentiation, Quantitative RT-PCR, Two Tailed Test

Journal: Brazilian Journal of Cardiovascular Surgery

Article Title: Expression Level and Significance of Tim-3 in CD4 + T Lymphocytes in Peripheral Blood of Patients with Coronary Heart Disease

doi: 10.21470/1678-9741-2020-0509

Figure Lengend Snippet: Upregulation of the expression of IL-7 in peripheral blood serum of patients with CHD. The expression level of IL-7 in peripheral blood serum of the study subjects in each group was detected by ELISA. *Compared with the control group, *P<0.05, **P<0.01; # P<0.05.

Article Snippet: The level of IL-7 in peripheral blood serum was detected using an IL-7 ELISA kit (SEKH-0015, Solarbio, Beijing, China) in strict accordance with the instructions of the kit.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Control

Journal: Brazilian Journal of Cardiovascular Surgery

Article Title: Expression Level and Significance of Tim-3 in CD4 + T Lymphocytes in Peripheral Blood of Patients with Coronary Heart Disease

doi: 10.21470/1678-9741-2020-0509

Figure Lengend Snippet:

Article Snippet: The level of IL-7 in peripheral blood serum was detected using an IL-7 ELISA kit (SEKH-0015, Solarbio, Beijing, China) in strict accordance with the instructions of the kit.

Techniques: Enzyme-linked Immunosorbent Assay, Polymerase Chain Reaction, Reverse Transcription, Standard Deviation

Journal: Nature immunology

Article Title: Microbial ligand-independent regulation of lymphopoiesis by NOD1

doi: 10.1038/s41590-023-01668-x

Figure Lengend Snippet: a , Total cell numbers in BM, thymus, spleen and lymph nodes from WT ( n = 12), NOD1 −/− ( n = 10), ΔCARD NOD1 ( n = 10), NOD2 −/− ( n = 6) or MyD88 −/− ( n = 6) mice. b , Absolute numbers of T and B lymphocytes and NK cells in spleen from WT, NOD1 −/− or ΔCARD NOD1 ( n = 10) mice. c , Level of IL-7, Flt3L, thrombopoietin (ThPO) or SCF measured in sera of individual WT, NOD1 −/− and ΔCARD NOD1 ( n = 13) mice by ELISA. d , Total numbers of LT-HSCs (Lin neg IL-7Rα neg Sca-1 + c-kit + Flt3 neg CD34 neg ), short-term HSCs (ST-HSCs) (Lin neg IL-7Rα neg Sca-1 + c-kit + Flt3 neg CD34 + ), CLPs (Lin neg IL-7Rα + Flt3 + Sca1 inter c-kit inter ) and common myeloid progenitors (CMPs) (Lin neg IL-7Rα neg Sca-1 neg c-kit + CD16/32 neg CD34 + ) in the BM of WT, NOD1 −/− or ΔCARD NOD1 ( n = 16) mice. e , Frequency of donor cells in BM, thymus and spleen 8 weeks after reconstitution of irradiated H-2K/D d/b WT recipient mice ( n = 5) with a combination of equal numbers of congenically marked sort-purified HSCs (Lin neg IL-7Rα neg c-kit + Sca-1 + Flt3 neg ) from H-2K/D b/b WT (CD45.1/2 + ), NOD1 −/− (CD45.2 + ) and ΔCARD NOD1 (CD45.1 + ) animals as indicated on the diagram. a – d , Each symbol represents the value for an individual mouse. The bars indicate mean ± s.e.m. value for each group pooled from three experiments ( a – c ) and from one representative out of two independent experiments performed ( d ). Significance was determined by two-sided Mann–Whitney test in a , b and e and one-way ANOVA (Tukey’s multiple comparisons) test in c and d and are indicated by asterisks: * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The hematopoietic cytokines were measured in sera of individual mice using Quantikine ELISA kits (R&D Systems: no. m700 for IL-7, no. MFK00 for Flt3L, no. MTP00 for thrombopoietin and no. MCK00 for SCF).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Irradiation, Purification, MANN-WHITNEY

Journal: Cell stem cell

Article Title: Bone marrow myeloid cells regulate myeloid-biased hematopoietic stem cells via a histamine-dependent feedback loop

doi: 10.1016/j.stem.2017.11.003

Figure Lengend Snippet: (A) TLR4 mRNA in HSCs (n = 3 - 4) and myeloid cells (n = 5). (B) TLRs expression from gene microarray analysis in myeloid cells (n = 3 per group). (C) Quantification of HSCs, HSPCs, and myeloid cells at 24 hours after LPS treatment (n = 5). (D) Myeloid cells (Gr1+) and progenitors (c-kit+) in spleen from LPS (n = 4) or PBS (n = 3)-treated Hdc-GFP mice. Four independent experiments. (E and F) Cell cycle analysis of BM HSCs at 6 hours after PBS (n = 3) or LPS (n = 3) treatment. (G) Absolute numbers of HSC in BM of Hdc-GFP mice treated with PBS, IFN-γ, or IL6 (n = 5 per group). Data were analyzed with two-tailed Student’s t-test (A, F, and G) or one-way analysis of variation (ANOVA) with Bonferroni post-hoc test (C). See also Figure S3.

Article Snippet: Proteins from serum or bone marrow supernatant were measured by using ELISA Kit (Histamine: Enzo Life Science; IL6, IL7, and IFN-γ: Thermo Fisher Scientific) according to manufacturer’s protocol.

Techniques: Expressing, Microarray, Cell Cycle Assay, Two Tailed Test

Journal: Cell stem cell

Article Title: Bone marrow myeloid cells regulate myeloid-biased hematopoietic stem cells via a histamine-dependent feedback loop

doi: 10.1016/j.stem.2017.11.003

Figure Lengend Snippet: (A) Relative mRNA expression of IL-7/IL7-R pathway genes in HSPCs (n = 4 per group). (B) Quantification of Hdc-GFPhi HSPCs in IL-7 or PBS-treated mice (n = 3 per group). (C) Percentage of G0 HSCs (n = 3 per group). (D) Frequencies of progenitors in IL-7 or PBS-treated Hdc-GFP mice (n = 5 per group). (E) Schematic depiction of the relative lineage bias of Hdc-GFPhi HSCs or Hdc-GFPlo HSCs. Data were analyzed with two-tailed Student’s t-test (A-D). See also Figure S3.

Article Snippet: Proteins from serum or bone marrow supernatant were measured by using ELISA Kit (Histamine: Enzo Life Science; IL6, IL7, and IFN-γ: Thermo Fisher Scientific) according to manufacturer’s protocol.

Techniques: Expressing, Two Tailed Test

Journal: Cell stem cell

Article Title: Bone marrow myeloid cells regulate myeloid-biased hematopoietic stem cells via a histamine-dependent feedback loop

doi: 10.1016/j.stem.2017.11.003

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Proteins from serum or bone marrow supernatant were measured by using ELISA Kit (Histamine: Enzo Life Science; IL6, IL7, and IFN-γ: Thermo Fisher Scientific) according to manufacturer’s protocol.

Techniques: Purification, Recombinant, Lysis, Staining, Enzyme-linked Immunosorbent Assay, Sequencing, Isolation, SYBR Green Assay, Expressing, Software

Journal: Cell & Bioscience

Article Title: S100A8/A9-amplified inflammation (TNF-α/IL-1β) in the bone marrow of tumor-bearing host disrupts early B cell development

doi: 10.1186/s13578-026-01565-4

Figure Lengend Snippet: Remote tumor-induced aberrant BM inflammation (TNF-α/IL-1β) disrupts CLP development. A Prediction of ligand-receptor interactions between CLPs (receiver) and other bone marrow cell populations (sender) in C57BL/6J tumor-bearing mice, analyzed by scRNA-seq data. B Measuring cytokines secretion changes in bone marrow fluid of control and tumor-bearing C57BL/6J mice (Lewis and B16F10) by ELISA. C Representative in situ IF staining of IL-7 expression in femur of control and tumor-bearing C57BL/6J mice (Scale bar: 20 μm) and IL-7 MFI was analyzed. D Measuring different cytokines secretion changes in blood of control and tumor-bearing C57BL/6J mice (Lewis) by Protein Microarray( N = 5). E Measuring CCL2, CCL4, G-CSF, TNF-α and IL-1β secretion changes in blood of control and tumor-bearing C57BL/6J mice (B16F10) by ELISA. F BM CLPs were isolated from healthy C57BL/6J mice and cultured in vitro for 5 days. Different cytokines were added to the culture system. Their effects on the differentiation ability of CLPs into B cells was assessed by microscopy and flow cytometry. G The number of B220 + cells were quantified. H Various cytokine concentrations of TNF-α or IL-1β were added to the culture system. Their effects on the differentiation ability of CLP into B cells was assessed by microscopy and flow cytometry. I The number of B220 + cells were quantified. J TNF-α and IL-1β were added simultaneously to the culture system, their effects on the differentiation ability of CLPs into B cells was assessed by microscopy and flow cytometry. K The number of B220 + cells were quantified. Data are representative of three independent experiments and were analyzed by two-tailed unpaired t-test. Two-tailed p-values were reported. Bar graphs denote mean values with SEM (* P < 0.05, ** P < 0.01 and *** P < 0.001)

Article Snippet: Mouse IFN-γ ELISA Kit and Mouse IL-7 ELISA Kit were from JINGMEI (Jiangsu, China).

Techniques: Control, Enzyme-linked Immunosorbent Assay, In Situ, Staining, Expressing, Microarray, Isolation, Cell Culture, In Vitro, Microscopy, Flow Cytometry, Two Tailed Test