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

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Copyright: © Yao et al.

This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.

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Figure 3. — Knockdown of NEAT1 reverses H/R-induced cell damage by reducing NLRP3 inflammasome activation-dependent pyroptosis. (A) The expression of NEAT1 mRNA was examined by RT-qPCR. Detection of (B) cell viability and (C) LDH activity in human umbilical vein endothelial cells following H/R. RT-qPCR analysis of (D) NEAT1, (E) miR-204 and (F) BRCC3 expression. (G) The expression levels of BRCC3, NLRP3, ASC, cleaved CASP1 and cleaved GSDMD protein were examined by western blotting. The expression levels of (H) IL-1β and (I) IL-18 mRNA were examined by RT-qPCR. The levels of secreted (J) IL-1β and (K) 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, micro RNA; RT-qPCR, reverse transcription-quantitative PCR; NC, negative control; CCK-8, Cell Counting Kit-8; shRNA, short hairpin RNA.