Fig. 5

Effects of low expression of BNIP3 on proliferation and apoptosis of RA-FLSs. A The effects of low expression of BNIP3 on the viability of RA-FLSs were detected by CCK-8 method (mean ± SD, n = 6 from three independent experiments); B BrdU was incorporated into RA-FLSs, cells were exposed to 21% and 2% O₂ for 24 h, and then BrdU (green) and DAPI (blue) immunostaining were detected; C Flowcytometry analysized the cell proliferation with PI staining after cells were treated with different oxygen concentrations for 24 h; D Histogram depicts quantitative data from the flow cytometric analysis (mean ± SD, n = 5 from three independent experiments); E RA-FLSs were analyzed by FCM using Annexin V/PI double-staining; F Histogram depicts the quantitative data from the AV/PI double-staining (mean ± SD, n = 6 from three independent experiments); G Representative fluorescence images of cells stained with TUNEL and Propidium Iodide to visualize apoptosis cells (Green) and nucleus (Blue); H The expressions of Bax and Bcl-XL was analyzed by Western blot (mean ± SD, n = 6 from three independent experiments). ^#P < 0.05, ^##P < 0.01 represents the comparison between Ctr siRNA N and BNIP3 siRNA N; *P < 0.05, **P < 0.01 represents the comparison between Ctr siRNA N and Ctr siRNA H, ⁺P < 0.05, ⁺⁺P < 0.01 represents the comparison between BNIP3 siRNA N and BNIP3 siRNA H, ^ΔP < 0.05, ^ΔΔP < 0.01 represents the comparison between Ctr siRNA H and BNIP3 siRNA H