Fig. 7

MiR-146a-5p targeting TRAF6 inhibits the differentiation of porcine intramuscular preadipocytes via the AKT/mTORC1 signaling pathway. a TRAF6 was predicted to be a target of miR-146a-5p by TargetScan software. b miR-146a-5p and TRAF6 3′ UTR region base complementary pattern diagram. c WT and MUT psiCHECK-2.0-TRAF6 vectors. d Relative luciferase activity of TRAF6 responding to miR-146a-5p mimics. To verify that miR-146a-5p can function by targeting TRAF6, we co-treated cells with miR-146a-5p mimics and TRAF6 overexpression vector (500 ng per hole, 6-well plate). e, f Bodipy or AdipoRed staining was performed on lipid droplets. g White light field and oil red O stained lipid droplets. h After extracting oil red O with isopropanol, the OD value (510 nm) was detected. i, j RT-qPCR was used to detect the TRAF6 and adipogenesis genes C/EBPβ, PPARγ, and FABP4. k Western blot analysis of TRAF6, C/EBPβ, PPARγ, and FABP4. l Protein quantitative analysis of k. m Western blot analysis of AKT, p-AKT, mTORC1, and p-mTORC1. n Protein quantitative analysis of m. Values are expressed as the mean ± SE (n = 3). *, P < 0.05; **, P < 0.01, versus NC