Figure 5

The effects of IL-1β stimulated hUCMSCs induced in HFLS-RA cells apoptosis and caspases activation. (a) After 24 h of co-culture, cells were stained with Annexin V/PI detection kit. hUCSMCs were labeled with CellTracker Orange. Annexin V represented as early apoptosis, and PI represented as late apoptosis. Green: Annexin V, red: nucleus-PI, whole cell with light red—hUCMSCs, scale bar: 100 μm. (b) Cleaved caspase-8, (e) cleaved caspase-9, and (h) cleaved caspase-3 expression in HFLS-RA cells and HFLS-RA cells co-cultured with hUCMSCs. The quantitative graphs of the Western blot results of (c,f,i). Full-length Western blots are in Supplementary Fig. S6. The data represent mean ± SD (n = 3) (*P < 0.05, **P < 0.01, ***P < 0.001). The expression of (d) cleaved caspase-8, (g) cleaved caspase-9, and (j) cleaved caspase-3 expression on HFLS-RA cells after co-culturing with hUCMSCs by immunofluorescence study. White blocks chosen are magnified in the column below. Orange: hUCMSCs labeled with CellTracker Orange, Green: cleaved caspase, blue: Hoechst 33258 (nucleus), scale bar: 100 μm.