Fig. 6

Effects of low expression of BNIP3 on the mitophagy and oxidation indexes of RA-FLSs. A RA-FLSs were analysed by electron microscopy. B RA-FLSs were infected with EGFP-LC3 (Green) for 24 h and then exposed to 21% and 2% O₂ for another 24 h, after that, mitochondria were marked with Mitotracker (Red). Representative images are shown. C The expression of Beclin-1 and LC3 was analyzed by Western blot (mean ± SD, n = 6 from three independent experiments); D RA-FLSs were exposed to 21% O₂ or 2% O₂ for 24 h and assessed the ROS content by FCM using 10 μM CM-H2DCFDA as a ROS probe; EC-1 was used to detect the effect of hypoxia on MMP of RA-FLSs; F Histogram depicts the quantitative data from the JC-1 staining (mean ± SD, n = 6 from three independent experiments); G The expressions of HIF-1α and BNIP3 in RA-FLSs after hypoxia for 24 h were 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