Fig 2: Etanercept attenuated the apoptosis of CSE-induced HPAECs via regulating levels of cell apoptosis-related proteins. (A–F) Protein levels of Bax (C), Bcl-2 (D), FasL (E) and Fas (F) after CSE exposure and ETN treatment were measured by Western blot. ß-Actin was used as the internal control. All experiments were performed in triplicate and experimental data were expressed as mean ± standard errors (SE). *P<0.05, vs control; #P<0.05, vs CSE.

Fig 3: Effect of AIV on the apoptosis of RPE cells from rats with DM. (A) AIV significantly decreased the apoptosis of RPE cells from rats with DM as shown by TUNEL assay, (a) Sham, (b) DM, (c) DM+H-AIV, (d) percentage apoptotic cells; scale bar, 100 µm; (B) representative apoptosis-related protein expression bands in RPE cells from rats with DM; (C) statistical analysis of gray value of apoptosis-related protein bands, (a) Bcl-2, (b) Bax, (c) Bcl-2/Bax, (d) Fas, (e) FasL, (f) Fas/FasL, (g) active caspase-3, (h) active caspase-8, (i) active caspase-9. *P<0.05 and ***P<0.001 vs. DM group, and ###P<0.001 vs. Sham group. Data are expressed as the means ± SD of 3 individual experiments. AIV, astragaloside-IV; DM, diabetes mellitus; RPE cells, retinal pigment epithelial cells.

Fig 4: Blockade of FasL or CX3CR1 at different phases of AIA-attenuated joint inflammation.(A–F) Ankle thicknesses of WT mice injected with anti-FasL or anti-CX3CR1 antibodies (10 µg per mouse daily) on days 0–9 (A, D), days 5–9 (B, E), and days 8–10 (C, F) to measure AIA (A–C, n = 6; D–F, n = 4). The arrows in the diagrams indicate the day on which antibodies were first injected. Data were pooled from three independent experiments and are presented as mean ± SEM. *p<0.05, **p<0.01, ***p<0.005. Data were analyzed using one-way ANOVA.Figure 6—source data 1.Numerical data obtained during experiments represented in Figure 6, Figure 6—figure supplement 1.

Fig 5: FasL and TRAIL compete for DR5 binding and exert similar effects on cell death.(A, B) FasL–Fc binding to hFLSCs or EL4 cells transfected with human FAS, or TNFRSF10B after preincubation with human sTRAIL or sFasL. (C) Model of FasL and DR5 derived from the crystal structure of the FasL/DcR3 complex (Protein Data Bank: 4 MSV) and TRAIL/DR5 complex (Protein Data Bank: 1D4V). (D, E) Flow cytometric analysis of FasL–Fc or DR5–Fc binding to EL4 cells transfected with human WT or mutated TNFRSF10B or FASLG. (F, G) Comparison of the effects of sFasL and sTRAIL on (F) apoptosis and (G) necroptosis in hFLSCs. (H) Joint swelling and clinical scores in Faslgld/gld mice injected with Z–VAD–FMK and/or sFasL (n = 6 per group). Data were pooled from four (A, B, and D–G) or three (H) independent experiments and are presented as mean ± SEM. *p<0.05; **p<0.01; ***p<0.005. Data were analyzed using one-way ANOVA.Figure 2—source data 1.Numerical data obtained during experiments represented in Figure 2, Figure 2—figure supplement 3.