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. 2020 Jul 23;22(4):2695–2704. doi: 10.3892/mmr.2020.11351

Figure 5.

Figure 5.

Silencing miR-223-3p reverses the effect of R.I flagellin on NLRP3 inflammasome activation and pyroptosis in THP-1 macrophages. (A) RT-qPCR analysis was performed to determine the expression levels of miR-223-3p, miR-5197-3p, miR-589-3p and miR-1305. (B) Schematic diagram of the predicted binding sequence of miR-223-3p in the 3′-UTR of NLRP3. (C) Luciferase activity for the WT and MUT 3′-UTR of NLRP3 pGL3 vector co-transfected with miR-223-3p NC, miR-223-3p mimics, miR-223-3p inhibitor NC and miR-223-3p inhibitor. RT-qPCR analysis was performed to determine the expression levels of (D) miR-223-3p and (E) NLRP3. (F and G) Western blot analysis was performed to determine NLRP3 protein expression. (H) IL-1β levels in the culture medium were detected via the enzyme-linked immunosorbent assay. (I) Western blot analysis was performed to determine the protein expression levels of (J) cleaved caspase-1 and (K) GSDMD-N. Data are presented as the mean ± standard error of the mean (n=3/group). *P<0.05. miR/miRNA, microRNA; R.I, Roseburia intestinalis; NLRP3, nucleotide-binding oligomerization segment-like receptor family 3; RT-qPCR, reverse transcription-quantitative PCR; IL, interleukin; GSDMD-N, Gasdermin D N-terminal fragment; n.s, no significance; UTR, untranslated region; NC, negative control; WT, wild-type; MUT, mutant.