Research Paper Volume 14, Issue 15 pp 6006—6027

Figure 4. Senolytics (ABT263 and D+Q) decrease the senescence phenotype population and axon guidance and angiogenic factors in fibroblast-like synovial cells from human OA tissue. Quantification of (A) SA-β-gal+ (n = 6 per group) and (B) TUNEL+ cells in human OA fibroblast-like synovial cells treated with ABT263 (2.5 μM, 1.25 μM, and 0.625 μM) (n = 3 per group). Representative images of (C) SA-β-gal staining and (D) TUNEL staining of synovial cells treated with 1.25 μM of ABT263 or Veh for 3 days. (E) Relative mRNA expression levels of CDKN2A and CDKN1A in the synovial cells treated with ABT263 (1.25 μM) or Veh (n = 5 for Veh, n = 6 for ABT263). Percentage of (F) SA-β-gal + (n = 6 per group) and (G) TUNEL+ cells (n = 3 per group) in D+Q (D 1000 nM, 500 nM, 250 nM + Q 200 μM, 100 μM, or 50 μM). Representative images of (H) SA-β-gal staining and (I) TUNEL staining of human OA synovial cells treated with D+Q (D 500 nM + Q 100 μM) for 3 days. (J) Relative mRNA expression levels of CDKN2A and CDKN1A in the synovial cells treated with D+Q (D 500 nM + Q 100 μM) or Veh (n = 5 per group). Relative mRNA expression levels of NGF, VEGF, and ATF4 in the synovial cells treated with (K) ABT263 (1.25 μM), Veh (n = 5 for Veh, n = 6 for ABT263), (L) D+Q (D 500 nM + Q 100 μM), or Veh (n = 5 for Veh, n = 6 for D+Q). Whisker plots represent the 10^th and 90^th percentiles, and the line corresponds to the median. Two independent experiments were performed. * p < 0.05, ** p < 0.01, *** p < 0.001; Unpaired Student’s t-test. Scale bars are shown in each image.