Overexpression of lncRNA GAS5 induces characteristic features of pyroptosis in MRC-5 cells. (A) Protein expression levels of pro-caspase-1, caspase-1, IL-1β, IL-18 and N-GSDMD in GAS5-overexpressing MRC-5 cells were measured using western blotting and three sets of representative blots are shown. (B) Semi-quantification of gene expression normalized to GAPDH. (C) Immunofluorescence staining showing the levels of caspase-1 in GAS5-overexpressing MRC-5 cells. Scale bar, 50 µm. (D) Quantitative measurement of IL-2, IL-6, IL-10 and TNF-α in GAS5-overexpressing MRC-5 cells using ELISA. (E) Protein expression levels of NLRP3 in GAS5-overexpressing MRC-5 cells were measured using western blotting. Semi-quantification of NLRP3 expression normalized to GAPDH and three sets of representative blots are shown. The results are presented as the mean ± SD (n=3; *P<0.05, **P<0.01, ***P<0.001 vs. control). N-GSDMD, cleaved N-terminal gasdermin D; GAS5, growth arrest-specific 5; lncRNA, long non-coding RNA; NLRP3, NLR family pyrin domain containing 3; NC, negative control; OE, overexpression.