Journal: Stem Cell Research & Therapy

Article Title: Bone marrow mesenchymal stem cells alleviate liver fibrosis after rat liver transplantation through JAK1/STAT5 pathway

doi: 10.1186/s13287-025-04353-y

Figure Lengend Snippet: BMSCs may alleviate liver fibrosis after liver transplantation through the JAK1/STAT5 pathway. ( A ) Box plot for FPKM values. PCA analysis. Bar plot of the statistic of differentially expressed genes. Venn graph of the differentially expressed genes. Volcano plot of the differentially expressed genes. Heat map of RNA differential expression. GO analysis reveals the potential impact of the differentially expressed RNAs on functions. KEGG analysis reveals the potential impact of the differentially expressed RNA on pathways. KEGG map of the JAK/STAT pathway. ( B ) Western blot analyses of JAK1, p-STAT5, STAT5, p-STAT1, STAT1, p-STAT3, STAT3, IL7R, and IL7 in liver tissue. TUBA1A was used as the loading control except for p-STAT5, p-STAT1, and p-STAT3. STAT5, STAT1, and STAT3 were used as the loading control for p-STAT5, p-STAT1, and p-STAT3, respectively. Jak1 , Il7r , and Il7 mRNA levels in liver tissue. ( C ) Microscopic images of each group stained with HE, Masson, Sirius Red, and ACTA2 (magnification: ×100, n = 6). Area stained with Masson, Sirius Red, and ACTA2, as well as Metavir fibrosis staging. The levels of serum ALT, AST, and TBil in each group. ( D ) Western blot analyses of COL1A1, JAK1, p-STAT5, STAT5, ACTA2, and CASP3 in liver tissues. TUBA1A was used as the loading control except for p-STAT5. STAT5 was used as the loading control for p-STAT5. Full-length blots/gels are presented in Supplementary Fig. . The samples were derived from the same experiment and processed in parallel. Col1a1 , Jak1 , Acta2 , and Casp3 mRNA levels in liver tissues. ACTA2, actin alpha 2; ALT, alanine aminotransferase; AST, aspartate aminotransferase; BMSCs, bone mesenchymal stem cells; CASP3, caspase 3; COL1A1, collagen type I alpha 1 chain; GO, Gene Ontology; IL, interleukin; JAK, Janus-activated kinase; KEGG, Kyoto Encyclopedia of Genes and Genomes; LT, liver transplantation; STAT, signal transducer and activator of transcription; TBil, total bilirubin. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: The LT + anti-IL7 group received anti-IL7 antibody (M25; Cat# HY-P990210, MCE, Shanghai, China) (250 μg/kg of body weight, per ip, once a week) for 2 weeks after LT.

Techniques: Transplantation Assay, Quantitative Proteomics, Western Blot, Control, Staining, Derivative Assay

Journal: Stem Cell Research & Therapy

Article Title: Bone marrow mesenchymal stem cells alleviate liver fibrosis after rat liver transplantation through JAK1/STAT5 pathway

doi: 10.1186/s13287-025-04353-y

Figure Lengend Snippet: BMSCs may inhibit the activation of HSC-T6 cells after hypoxia–reoxygenation and reduce their apoptosis through the JAK1/STAT5 pathway. ( A ) Western blot analyses of JAK1, p-STAT5, STAT5, p-STAT1, STAT1, p-STAT3, STAT3, IL7R, and IL7 in HSC-T6 cells. TUBA1A was used as the loading control except for p-STAT5, p-STAT1, and p-STAT3. STAT5, STAT1, and STAT3 were used as the loading control for p-STAT5, p-STAT1, and p-STAT3, respectively. Jak1 and Il7r mRNA levels in HSC-T6 cells. ( B ) Western blot analyses of COL1A1, JAK1, p-STAT5, STAT5, and ACTA2 in HSC-T6 cells. TUBA1A was used as the loading control except for p-STAT5. STAT5 was used as the loading control for p-STAT5. Col1a1 , Jak1 and Acta2 mRNA levels in HSC- T6 cells. ( C ) Apoptosis results of flow cytometry in each group. Full-length blots/gels are presented in Supplementary Fig. . The samples were derived from the same experiment and processed in parallel. ACTA2, actin alpha 2; BMSCs, bone mesenchymal stem cells; COL1A1, collagen type I alpha 1 chain; IL, interleukin; JAK, Janus-activated kinase; STAT, signal transducer and activator of transcription. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: The LT + anti-IL7 group received anti-IL7 antibody (M25; Cat# HY-P990210, MCE, Shanghai, China) (250 μg/kg of body weight, per ip, once a week) for 2 weeks after LT.

Techniques: Activation Assay, Western Blot, Control, Flow Cytometry, Derivative Assay

Journal: Stem Cell Research & Therapy

Article Title: Bone marrow mesenchymal stem cells alleviate liver fibrosis after rat liver transplantation through JAK1/STAT5 pathway

doi: 10.1186/s13287-025-04353-y

Figure Lengend Snippet: BMSCs regulate the IL7R/JAK1/STAT5 pathway in HSCs by modulating the secretion of IL7 by hepatic cells. ( A ) Microscopic images of each group stained with HE, Masson, Sirius Red, ACTA2, p-STAT5 and STAT5 (magnification: ×100, n = 6). Area stained with Masson, Sirius Red, ACTA2, and p-STAT5/STAT5, as well as Metavir fibrosis staging. The levels of serum ALT, AST, and TBil in each group. ( B ) Western blot analyses of COL1A1, JAK1, p-STAT5, STAT5 and ACTA2 in liver tissue. TUBA1A was used as the loading control except for p-STAT5. STAT5 was used as the loading control for p-STAT5. Col1a1 , Jak1 , and Acta2 mRNA levels in liver tissue. ( C ) Western blot analyses of IL7 in IAR20 cells. TUBA1A was used as the loading control. Il7 mRNA levels in IAR20 cells. ( D ) Western blot analyses of COL1A1, JAK1, p-STAT5, STAT5 and ACTA2 in HSC-T6 cells. TUBA1A was used as the loading control except for p-STAT5. STAT5 was used as the loading control for p-STAT5. Col1a1 , Jak1 and Acta2 mRNA levels in HSC- T6 cells. Full-length blots/gels are presented in Supplementary Fig. . The samples were derived from the same experiment and processed in parallel. ACTA2, actin alpha 2; ALT, alanine aminotransferase; AST, aspartate aminotransferase; BMSCs, bone mesenchymal stem cells; COL1A1, collagen type I alpha 1 chain; IL, interleukin; JAK, Janus-activated kinase; LT, liver transplantation; STAT, signal transducer and activator of transcription; TBil, total bilirubin. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: The LT + anti-IL7 group received anti-IL7 antibody (M25; Cat# HY-P990210, MCE, Shanghai, China) (250 μg/kg of body weight, per ip, once a week) for 2 weeks after LT.

Techniques: Staining, Western Blot, Control, Derivative Assay, Transplantation Assay

Journal:

Article Title: Murine lymph node-derived stromal cells effectively support survival but induce no activation/proliferation of peripheral resting T cells in vitro

doi: 10.1046/j.1365-2567.2003.01693.x

Figure Lengend Snippet: Effects of known cytokines to support the survival of T cells. T cells isolated from lymph nodes of C57BL/6 mice were cultured with various concentrations (as indicated in the figure, per ml) of IL-4, IL-6, IL-7, and IFN-β for 24 or 48 hr, and cell viability was determined. The data of cells cultured in the stromal-conditioned medium (SCM) containing 50% of culture supernatant of the LNS cells were added in each panel to compare the effect of each cytokine with that of SCM. Three different experiments were performed, and the average values are shown.

Article Snippet: ELISA assay for IL-7 was performed using the above-mentioned ELISA set with recombinant murine IL-7, anti-mouse IL-7 antibody (R & D systems, Minneapolis, MN) and biotynilated anti-mouse IL-7 antibody (BD-PharMingen).

Techniques: Isolation, Cell Culture

Journal:

Article Title: Murine lymph node-derived stromal cells effectively support survival but induce no activation/proliferation of peripheral resting T cells in vitro

doi: 10.1046/j.1365-2567.2003.01693.x

Figure Lengend Snippet: Blocking of the activity to support T-cell survival with neutralizing antibodies against various cytokines. (a) T cells were cultured with 1·0 ng/ml (IL-4, IL-6 and IL-7) or 10 U/ml (IFN-β) of cytokines with or without indicated neutralizing antibodies (5 µg/ml) for 24 or 48 hr, and cell viability was determined. T cells were also cocultured with the confluent layer of LN-derived stromal cells (+ LNS) or cultured in SCM with or without a cocktail of all antibodies (5 µg/ml each) for 24 or 48 hr, and cell viability was determined. Three experiments were performed, and the data are means ± SE. (b) IL-4, IL-6, and IL-7 levels in SCM were measured by ELISA. Three independently prepared SCM were analysed. *Results below the reliability level in the ELISA system we used (<15·6 pg/ml).

Article Snippet: ELISA assay for IL-7 was performed using the above-mentioned ELISA set with recombinant murine IL-7, anti-mouse IL-7 antibody (R & D systems, Minneapolis, MN) and biotynilated anti-mouse IL-7 antibody (BD-PharMingen).

Techniques: Blocking Assay, Activity Assay, Cell Culture, Derivative Assay, Enzyme-linked Immunosorbent Assay

Journal:

Article Title: Murine lymph node-derived stromal cells effectively support survival but induce no activation/proliferation of peripheral resting T cells in vitro

doi: 10.1046/j.1365-2567.2003.01693.x

Figure Lengend Snippet: Cell surface markers of T cells cultured with LNS cells. Freshly isolated T cells and T cells cocultured with LNS cells, cultured in SCM or cultured with recombinant IL-4 (1 ng/ml), IL-7 (2 ng/ml), or IFN-β (100 U/ml) for 48 hr were stained with fluorescence-conjugated antibodies and analysed by flow cytometry. Numbers in each panel indicates the percentages of cells in a quadrant where the numbers are.

Article Snippet: ELISA assay for IL-7 was performed using the above-mentioned ELISA set with recombinant murine IL-7, anti-mouse IL-7 antibody (R & D systems, Minneapolis, MN) and biotynilated anti-mouse IL-7 antibody (BD-PharMingen).

Techniques: Cell Culture, Isolation, Recombinant, Staining, Fluorescence, Flow Cytometry

Journal:

Article Title: Murine lymph node-derived stromal cells effectively support survival but induce no activation/proliferation of peripheral resting T cells in vitro

doi: 10.1046/j.1365-2567.2003.01693.x

Figure Lengend Snippet: DNA synthesis and cell cycle analysis of T cells cocultured with lymph node-derived stromal (LNS) cells, cultured in SCM or cultured with a cytokine. (a) T cells isolated from C57BL/6 lymph nodes were cultured in a simple medium (SM), cocultured with LNS cells, cultured in SCM or cultured with the indicated recombinant cytokine. Concentrations of cytokines were 10 ng/ml for IL-4, IL-6 and IL-7 and 100 U/ml for IFN-β. After 24 or 48 hr of culture, cells were pulsed with [3H]thymidine, and its uptake was measured as described in Materials and Methods. Three independent experiments were performed, and data are presented as means ± SE. (b) Fresh T cells, T cells cultured with IL-7 (10 ng/ml) for 48 hr, T cells cultured in SCM, and T cells cultured with LNS cells for 48 hr were prepared, and the cell cycles were analysed as described in Materials and Methods. Markers and numbers in each panel represent the areas of G0/G1, S, and G2/M phases and the percentages of cells in the area, respectively.

Article Snippet: ELISA assay for IL-7 was performed using the above-mentioned ELISA set with recombinant murine IL-7, anti-mouse IL-7 antibody (R & D systems, Minneapolis, MN) and biotynilated anti-mouse IL-7 antibody (BD-PharMingen).

Techniques: DNA Synthesis, Cell Cycle Assay, Derivative Assay, Cell Culture, Isolation, Recombinant

Journal:

Article Title: Murine lymph node-derived stromal cells effectively support survival but induce no activation/proliferation of peripheral resting T cells in vitro

doi: 10.1046/j.1365-2567.2003.01693.x

Figure Lengend Snippet: Expression of Bcl family proteins in T cells cultured with LNS cells. Fresh T cells or T cells cultured with LNS cells or with a recombinant cytokine (IL-4, IL-6, and IL-7, 10 ng/ml) in the absence of supporting cells and cytokines (T alone) for 48 hr, were collected and analysed for their expression of Bcl-2 and Bcl-XL by immunoblotting as described in Materials and Methods. In the right panel, T cells were cultured with IFN-β (100 U/ml) for 24 hr with or without a neutralizing antibody and analysed for Bcl-XL expression.

Article Snippet: ELISA assay for IL-7 was performed using the above-mentioned ELISA set with recombinant murine IL-7, anti-mouse IL-7 antibody (R & D systems, Minneapolis, MN) and biotynilated anti-mouse IL-7 antibody (BD-PharMingen).

Techniques: Expressing, Cell Culture, Recombinant, Western Blot