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. 2021 Oct 15;13(10):11384–11398.

Figure 4.

Figure 4

MiR-22-3p which is the target of LncRNA H19, targets NLRP3. A: A subcellular fractionation analysis was performed to determine the localization of LncRNA H19 in the WI-38 cells; B: Bioinformatics analysis indicated miR-22-3p is the target of LncRNA H19 and NLRP3; C: RT-qPCR was carried our to assess the levels of the miRNAs in the LPS induced WI-38 cells, including miR-22-3p, miR-20a-5p, miR-106b-5p, miR-20b-5p, and miR-17-5p; *P<0.05, **P<0.01, vs. 0 (LPS μg/mL). D: TargetScan software was used to predict the potential binding site between LncRNA H19 and miR-22-3p. E: RT-qPCR was conducted to assess the miR-22-3p expression; **P<0.01, vs. NC mimic+LPS. F: A dual luciferase reporter gene assay was performed to confirm the direct binding relationship between LncRNA H19 and miR-22-3p. **P<0.01, vs. NC mimic. G: RT-qPCR was performed to assess the miR-22-3p expression; **P<0.01, LPS vs. the control; ##P<0.01, Si-LncRNA H19+LPS vs. Si-NC+LPS. H: TargetScan software was used to predict the potential binding site between miR-22-3p and NLRP3. I: A dual luciferase reporter gene assay was conducted to confirm the direct binding relationship between miR-22-3p and NLRP3. **P<0.01, vs. the NC mimic. J: RT-qPCR was carried out to assess the NLRP3 mRNA levels. **P<0.01, LPS vs. the control; ##P<0.01, miR-22 mimic+LPS vs. the NC mimic+LPS. K: Western blot was performed to measure the NLRP3 protein expression. **P<0.01, LPS vs. the control; ##P<0.01, miR-22 mimic+LPS vs. NC mimic+LPS. L, M: The pyroptosis-related gene levels were measured using RT-qPCR and Western blot, including GSDMD, GSDMD-N, NLRP3, ASC-1 and Caspase-1. *P<0.05, **P<0.01, Si-LncRNA H19+miR-22-3p inhibitor+LPS vs. Si-LncRNA H19+NC inhibitor+LPS.