Fig. 3

Effect of H₂O₂ on mitochondrial dysfunction, ER stress, and AMPK signaling in BMSCs. BMSCs were treated with H₂O₂ at different concentrations from 200 to 800 μM for 24 h, and then (a) ROS production, (b) mitochondrial superoxide levels, and (c) MMP levels were analyzed. d Western blot measurement and quantification of Cyto-C translocation immunostaining in BMSCs after H₂O₂ injury for 24 h. e Representative images of Bip (red) and CRT (green) immunostaining in BMSCs treated with H₂O₂ for 24 h. Nuclei are labeled with DAPI. f After treatment with H₂O₂, the levels of p-AMPK and proteins related to ER stress, such as p-PERK, Bip, p-eIF2α, and DDIT3, were determined using western blotting in BMSCs, with normalization to the values of β-actin. Cells treated with PBS served as a control group. Values are expressed as the mean ± SD (n = 3 independent experiments). ^*p < 0.05 compared with the control group. One-way ANOVA followed by the LSD test was used to analyze the significance of differences. Scale bar = 50 μm. CRT, calreticulin; Cyto-C, Cytochrome C; ROS, reactive oxygen species