Fig. 2

NLRP3 inflammasome is activated and induces pyroptosis after radiation of lung epithelial cells. A qRT-PCR showing that the mRNA level of NLRP3, CASP1, IL-1β and GSDMD expression were increased after radiation in BEAS-2B (n = 3). B Western blotting showing that the protein level of NLRP3, CASP1 cleavage and GSDMD cleavage were increased after radiation in BEAS-2B (n = 3). C Representative fluorescent microscopy images of BEAS-2B that was co-stained for NLRP3, ASC and DAPI before and after radiation (n = 3). Scale bar, 50 μm. D ELISA results of IL-1β in supernatant of BEAS-2B, H446, A549 and H460 at different time points after radiation (n = 3). E BEAS-2B cells were transfected with the control or siNLRP3 siRNA, and given 10 Gy X-ray radiation, then samples were collected and changes in essential components of NLRP3 inflammasome were detected (n = 3). F ELISA results of IL-1β in supernatant of BEAS-2B transfected with control or NLRP3 siRNA at different time point after radiation (n = 3). G Microscopy images of PHBE and fluorescent microscopy images of PHBE stained for epithelium-specific cytokeratins and fibroblast marker α-SMA (n = 3). H Western blotting showing the level of NLRP3 protein expression, CASP1 cleavage and GSDMD cleavage were increased after radiation in PHBE (n = 3). I ELISA results of IL-1β in supernatant of PHBE 48 h after radiation (n = 3). Bar graphs show the means ± SEM; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. qRT-PCR quantitative reverse transcription polymerase chain reaction, ASC apoptosis-associated speck-like protein containing a caspase-recruitment domain, IL interleukin, ELISA enzyme-linked immunosorbent assay, PHBE primary human bronchial epithelial cells, SEM standard error of mean