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. 2021 Jan 26;11:616141. doi: 10.3389/fimmu.2020.616141

Figure 2.

Figure 2

The miR-223-3p is a negative regulator for S1pr1. (A) HUVECs were co-transfected with nontargeting miRNA and miR-223-3p mimic with the no UTR or 3′UTR. Luciferase activity was assayed 24 h after transient co-transfection. Renilla luciferase activity was measured. (n = 3 per group). Data are presented as mean ± SEM. *p < 0.05, ns, not significant, by Wilcoxon signed-rank test. (B) The mRNA and protein level of S1pr1 in EL4 cells was analyzed by TaqMan quantitative PCR and Western blot analyses after transfection with miR-223-3p mimic or nontargeting miRNA. The transfection efficacy of miR-223-3p mimic was confirmed by TaqMan quantitative PCR. miR-223-3p was transfected by lipofection, and the expression was compared with the nontargeting miRNA after 24 h transfection. The mRNA and protein levels of S1pr1 after the overexpression of miR-223-3p was confirmed by TaqMan quantitative PCR and Western blot analyses. Numbers above the Western blots indicate band intensity (normalized to total GAPDH) measured by using ImageJ software. Quantification of Western blot results (right side). (n = 3 per group). Data are presented as mean ± SEM. *p < 0.05, ns, not significant, by Wilcoxon signed-rank test.