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. 2021 Nov 29;25(1):32. doi: 10.3892/mmr.2021.12548

Figure 5.

Figure 5.

NEAT1 aggravates H/R-induced cell damage and NLRP3 inflammasome activation-dependent pyroptosis and is rescued by miR-204 overexpression. HUVECs were co-transfected with miR-204 or miR-con and pcDNA-NEAT1 or pcDNA-con before H/R. Detection of (A) cell viability and (B) LDH activity in HUVECs following H/R. RT-qPCR analysis of (C) NEAT1, (D) miR-204 and (E) BRCC3 expression. (F) The expression levels of BRCC3, NLRP3, ASC, cleaved CASP1 and cleaved GSDMD protein were examined by western blotting. The expression levels of (G) IL-1β and (H) IL-18 mRNA were examined by RT-qPCR. The levels of secreted (I) IL-1β and (J) IL-18 were evaluated by ELISA. Data are shown as the mean ± SD from five different experiments. **P<0.01. H/R, hypoxia/reoxygenation; LDH, lactate dehydrogenase; IL, interleukin; NLRP3, NOD-like receptor protein 3; ASC, apoptosis speck-like protein; CASP1, caspase-1; GSDMD, gasdermin D; NEAT1, nuclear enriched abundant transcript 1; BRCC3, BRCA1/BRCA2-containing complex subunit 3; miR, microRNA; RT-qPCR, reverse transcription-quantitative PCR; HUVEC, human umbilical vein endothelial cell; CCK-8, Cell Counting Kit-8.