Fig. 3
ENO1 induces PASMC resistance to apoptosis. a The amount of apoptotic markers: PARP (full length and cleaved), caspase 3 (full length and cleaved), caspase 9 (full length and cleaved) were detected by western blotting in shENO1–PASMC. b shENO1 and shCTRL PASMC were subjected to the TUNEL assay (Scale bars, 100 μm) and c the apoptotic rates were calculated (n = 3, **P < 0.0001) with the TUNEL assay in shENO1–PASMC. d PASMC were treated with ENOblock in different concentrations for 24 h, and the LDH assay was performed to detect cell death. e PASMCs were treated with 10 μM ENOblock and hypoxia (1% O2) for 24 h, and western blotting was used to detect apoptotic markers in cell lysate. f Images of the TUNEL assay (Scale bars, 100 μm) and g calculated apoptotic rates (n = 3 per group, **P < 0.0001) in ENOblock-treated PASMCs under normoxic and hypoxic conditions. h PASMCs were transfected with pCMV3-ENO1-GFP and treated with etoposide (in gradient concentrations) for 24 h. Afterwards, the cell viability was measured (n = 3 per group, *P < 0.05). i PASMCs were transfected with pCMV3-ENO1-GFP and treated with 100 μM etoposide for 24 h. The apoptotic markers were detected using western blotting in the cell lysate. j Representative images of the TUNEL assay were shown (Scale bars, 100 μm). k Apoptotic rates in ENO1-overexpressed PASMC under the treatment of etoposide were calculated from images in experiments depicted in j. n = 3 per group, *P = 0.04489. Data represent the mean ± SEM. Student t test and one-way ANOVA were used to compare two and multiple groups. Bonferroni post-tests were carried out after ANOVA
