Fig. 2.

Overexpression FENDRR inhibits hypoxia-induced HPAECs pyroptosis. a The protein levels of NLRP3 and Caspase-1 were elevated after hypoxia exposure 24 h (n = 6). b Overexpression FENDRR reversed the increased protein levels of Caspase-1, NLRP3, Pro-caspase-1 and IL-1β induced by hypoxia in HPAECs (n = 5). c The positive PI (red) cells were detected in hypoxic HPAECs transfected with FENDRR overexpression plasmid. Scale bar = 50 μm. d Fluorescence staining of Caspase-1 in HPAECs transfected with FENDRR overexpression plasmid. HPAECs were stained for Caspase-1 (red) and CD31 (green), and DAPI (blue) was used for nuclear staining. Scale bar = 50 μm. e The activity of caspase-1 was examined using caspase-1 activity assay kit (n = 5). f The Flow cytometry assay was used to detect the population of positive-PI cells. HPAECs were treated with annexin V-FITC/propidium iodide (PI) double staining using quantitative fluorescence-activated cell sorting (FACS) analysis. g HPAECs were transfected with FENDRR overexpression plasmid under hypoxia and the LDH release was evaluated by LDH release kit (n = 5). Each datapoint in the figure represents a unique biological replicate. All values are presented as the mean ± SD. Statistical analysis was performed with one-way ANOVA or the Student’s t-test. NOR: normoxic; HYP: hypoxic; NC: negative control. *P < 0.05, **P < 0.01, ***P < 0.001 compared with NOR + NC. ^#p < 0.05, ^##p < 0.01, ^###p < 0.001 compared with HYP + NC