Journal: The Journal of Clinical Investigation

Article Title: IL-17 and immunologically induced senescence regulate response to injury in osteoarthritis

doi: 10.1172/JCI134091

Figure Lengend Snippet: (A) Multiparametric flow cytometric analysis of CD8+, CD4+, and yδ+ T cells (CD45+CD3+) isolated from the joint compartment 1 week after sham surgery and ACLT compared with control mice with no surgery (N.S.) (n = 4). (B) Immunofluorescence of IL-17 in the synovium and cartilage 1 week after ACLT compared with no surgery in young mice. Scale bars: 25 μm. (C) Flow cytometric data showing IL-17a and IL-17f expression in ILCs from inguinal (Ig) LNs 4 weeks after ACLT (CD3–Thy1.2+NK1.1–). (D) Quantification of mRNA expression for inflammatory markers in ILCs (CD3–Thy1.2+) sorted from the joint compartment 2 weeks after ACLT (n = 2). (E) Percentage of Th17 cells in young (Y) and 18-month-old aged (A) animals 2 and 4 weeks after ACLT in the inguinal LNs, as determined by flow cytometry and immunofluorescence staining for CD4 and IL-17 in LNs from young mice 2 weeks after ACLT. Scale bar: 10 μm. (F) Quantification of Il17 mRNA expression in LN tissue (n = 3). (G) Quantification of Cdkn2a mRNA expression in young and aged animal joints with no surgery and in joints 2 and 4 weeks after ACLT (n = 3). (H) p16 staining of ACLT cartilage and no-surgery cartilage from young mice. Scale bars: 25 μm. Data indicate the mean ± SD. *P < 0.05, **P < 0.01, and ****P < 0.001, by 1-way ANOVA with Holm-Šidák multiple-comparisons test. All groups were compared with each other. (E–G) Separate 1-way ANOVAs were performed for each time point. F, femur; S, synovium; T, tibia; RQ, relative quantification.

Article Snippet: The Th17 control condition was made using the CellXVivo Human Th17 Differentiation Kit (R&D Systems).

Techniques: Isolation, Control, Immunofluorescence, Expressing, Flow Cytometry, Staining, Quantitative Proteomics

Journal: The Journal of Clinical Investigation

Article Title: IL-17 and immunologically induced senescence regulate response to injury in osteoarthritis

doi: 10.1172/JCI134091

Figure Lengend Snippet: (A) Percentage of mouse CD4+IL-17a+, CD4+IL-17f+, and CD4+IFN-γ+ T cells in coculture conditions: naive T cells alone (NT), SnC coculture, SnCs plus IL-2 plus TGF-β coculture, media with TGF-β (n = 3 for each group). Representative flow plots are shown. (B) Quantification of Cdkn2a and Il6 mRNA expression in fibroblasts cocultured with naive CD4+ T cells and Th17 cells (n = 3 for each group). Representative images of SA–β-gal staining of fibroblasts cocultured with naive CD4+ T cells and Th17 cells are shown. Scale bars: 25 μm. (C) Percentage of human CD4+IL-17a+ T cells after 7 days in coculture. Conditions from left to right: no coculture (NT), Th17 positive control, high percentage of SnCs coculture, high percentage of SnCs plus IL-2 plus TGF-β coculture, low percentage of SnCs coculture, low percentage of SnCs plus IL-2 plus TGF-β coculture (n = 3 for each group). (D) Volcano plot of Th17 cocultured cells normalized to normal fibroblasts. The dotted line denotes P = 0.05. Graph on the right indicates differentially regulated genes related to STAT3 and Wnt signaling. (E) Differential expression of the top-50 genes in immune-induced senescence (Th17 coculture) compared with irradiation-induced senescence, 7 days after senescence induction (n = 3). Data indicate the mean ± SD. *P < 0.05 and **P < 0.01, by 1-way ANOVA with Holm-Šidák multiple-comparisons test. All groups were compared with each other. A 2-tailed Student’s t test was performed within each time point in B.

Article Snippet: The Th17 control condition was made using the CellXVivo Human Th17 Differentiation Kit (R&D Systems).

Techniques: Expressing, Staining, Positive Control, Quantitative Proteomics, Irradiation