Figure 4.

miR-101 inhibits the expression of C/EBPα and KLF6

(A) The binding sites of miR-101-3p to KLF6 and C/EBPα. (B) Dual luciferase reporter assays were measured in the 293T cells cotransfected with the miR-101-mimic and luciferase reporter plasmid that was inserted with the wild-type (WT)/mutation (MT)-C/EBPα 3′ UTR or WT/MT-KLF6 3′ UTR. Luc, firefly luciferase. (C) Quantitative real-time PCR results showing the relative expression of miR-101 in M1 and M2 macrophages. (D−F) miR-101-mimic or NC-mimic was transfected into M1 macrophages for 24 h. Quantitative real-time PCR results showing the relative expression of miR-101 (D), quantitative real-time PCR results showing the relative mRNA expression of C/EBPα and KLF6 (E), and western blot results showing the protein expression of C/EBPα, KLF6, PU.1, and PPARγ (F). (G−I) miR-101-inhibitor or NC-inhibitor was transfected in M2 macrophages for 48 h. Quantitative real-time PCR results showing the relative expression of miR-101 (G), quantitative real-time PCR results showing the relative mRNA expression of C/EBPα and KLF6 (H), and western blot results showing the protein expression of C/EBPα, KLF6, PU.1, and PPARγ (I). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001; n = 3; mean ± SD.