Fig 2: Analysis of protein expression of the apoptosis biomarkers caspase 3 and Bax and the antiapoptosis indicator BCL-2 and a TUNEL assay in the urinary bladder at 4 days after IC induction (n = 5). a Expression of the apoptosis biomarkers caspase 3 and Bax and the antiapoptosis indicator BCL-2 by Western blot in the bladder tissue was highest in the interstitial cystitis (IC) group compared to the IC + urine-derived stem cell (USC) group and normal control (N) group; the biomarker expression in the IC + USC group was lower than in the IC group. b A statistical chart of the relative optical density of caspase 3/GAPDH in each group (n = 5). c A statistical chart of the relative optical density of Bax/GAPDH in each group (n = 5). d A statistical chart of the relative optical density of BCL-2/GAPDH in each group (n = 5). e–g The TUNEL assay indicated that, in sections from the IC group rats, the most apoptotic nuclei were observed, whereas fewer were present in the IC + USC group and more were present than in the normal control group. h A statistical chart reveals the index of apoptotic nuclei in all groups of bladder tissue (n = 5). *P < 0.05, versus the control group; # P < 0.05, versus the IC group

Fig 3: Nox hyperactivity contributes to cardiomyocyte apoptosis in SAP rats. Nox inhibition attenuates myocardial cell apoptosis and apoptosis-associated protein expression in the myocardium 24 h after SAP challenge. Representative images (400x magnification) of TUNEL assay (a). Immunoblot of Bax and Bcl-2 (b) and procaspase-3 and cleaved-caspase-3 (d) protein expressions from heart samples. Densitometry analysis of Bax and Bcl-2 (c) and procaspase-3 and cleaved-caspase-3 (e). Data are expressed as means ± SD, n = 3 fluorescence values per group; means ± SD, n = 3 immunoblot results per group. * p < 0.05 compared with the SO group. # p < 0.05 compared with the SAP group.

Fig 4: ALK7 promoted VSMC apoptosis via Smad2/3 signaling. (A–B). The total and phosphorylation of Smad2 and Smad3 in the aortas with atherosclerotic plaques of diabetic mice were determined by western blot. n = 3 per group. *p < 0.05 vs. the Chow group, # p < 0.05 vs. the DM + nc-shRNA group. (C–D). The total and phosphorylation of Smad2 and Smad3 in VSMCs co-treated with HG and PA were determined by western blot. n = 3 per group. *p < 0.05 vs. the control group, # p < 0.05 vs. the HG + PA + nc-shRNA group. (E–F). The protein level of ALK7, Bcl-2, Bax, and cleaved caspase 3 in VSMCs co-treated with HG and PA and Smad2/3 inhibitor with or without ALK7-shRNA were determined by western blot. n = 3 per group. *p < 0.05 vs. the control group, # p < 0.05 vs. the HG + PA group, & p < 0.05 vs. the HG + PA + inhibitor. Data was analyzed with ANOVA.

Fig 5: BIRF regulates GLT-1 expression and OGD tolerance in vitro (A and B) The effect of BIRF siRNA on the expression of GLT-1 mRNA by qRT-PCR (A) and protein by western blot (B) in astrocytes (n = 5). C, D Representative immunocytochemical staining photomicrographs show the effect of BIRF siRNA on GLT-1 expression in astrocyte (C) (scale bar = 50 µm) and its AOD measured using ImageJ software (D) (n = 3). E, F Hoechst-PI shows cell death of neurons (E) (scale bar = 50 µm) and statistical comparison of neuronal death rate (F) (n = 3). G CCK-8 assay was conducted to explore the influence of BIRF siRNA on cell viability of neurons (n = 7). H The effect of BIRF siRNA on caspase-3 (n = 8) and Bax/Bcl-2 (n = 5) expression in neurons by western blot. Data are presented as the mean ± SD. *P < 0.05, **P < 0.01