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. 2020 Oct 7;11:569173. doi: 10.3389/fimmu.2020.569173

Figure 5.

Figure 5

miR-302/367 cluster post-transcriptionally represses NF-κB expression by targeting its 3′UTR. (A) Sequences of mouse miR-302a and their predicted interactions with conserved miR-302a seeds found within the 3′UTRs of NF-κB are shown. The sequence of the NF-κB 3′UTR seed mutant used for the reporter assays and the predicted disruption of the miR-302a interaction are also shown. (B) Normalized luciferase activity of a reporter containing the wild-type or point-mutated 3′UTR reporter constructs (wt. UTR or mutant UTR) of NF-κB in MH-S cells co-transfected with control mimics or miR-302a and miR-302b mimics. (C, D) MH-S cells were transfected with miR-302a and miR-302b mimics (C) or inhibitors (D) for 24 h and then either left uninfected or infected with PA14 for 6 h. The expression levels of NF-κB mRNA were detected by qPCR (C, D). The expression levels of different cytokine mRNA were detected by qPCR (E). The medium supernatant was collected and different cytokines were measured by a standard ELISA (F–H). Data are shown as the mean ± SEM of three independent experiments. *p < 0.05; **p < 0.01; ***p < 0.001; NS, not significant.