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Fig. 4 | Journal of Animal Science and Biotechnology

Fig. 4

From: MiR-146a-5p targeting SMAD4 and TRAF6 inhibits adipogenensis through TGF-β and AKT/mTORC1 signal pathways in porcine intramuscular preadipocytes

Fig. 4

MiR-146a-5p targeting SMAD4 inhibits the proliferation of porcine intramuscular preadipocytes by the TGF-β signaling pathway. a SMAD4 was predicted to be a target of miR-146a-5p by TargetScan software. b miR-146a-5p and SMAD4 3′ UTR region base complementary pattern diagram. c WT and MUT psiCHECK-2.0-SMAD4 vectors. d Relative luciferase activity of SMAD4 responding to miR-146a-5p mimics. To verify that miR-146a-5p can function by targeting SMAD4, we co-treated cells with miR-146a-5p mimics and SMAD4 overexpression vector (500 ng, 6-well plate). e EdU staining assay. f Quantification ratio of EdU-positive cells/total cells. g CCK-8 analysis. h RT-qPCR was used to detect SMAD4 mRNA expression level. i Cell cycle genes cyclin B, cyclin D, cyclin E, and p21 expression level. j Western blot analysis of cyclin E, cyclin D, PCNA and p21. k Protein quantitative analysis of cyclin E, cyclin D, PCNA and p21. Values are expressed as the mean ± SEM (n = 3). *, P < 0.05; **, P < 0.01; ***, P < 0.001, versus NC

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