Fig. 3.

Loss of MyoD dramatically downregulates miR-133a, promotes brown adipocyte biogenesis and stimulates transdifferentiation of myoblasts into brown adipocytes. MyoD^iCre/+ mice were intercrossed to generate MyoD^iCre/iCre (MyoD^−/−) knockout mice and WT littermates. The miRNA miR-133a; Prdm16 targets miR-193b and miR-365; adipogenic marker miR-143 and brown adipogenic marker miR-455 were analyzed by qPCR. (a) qPCR analysis of WT and MyoD^−/− myoblasts for myogenic markers Myf5 and Myogenin, adipogenic markers Pparγ2 and Prdm16, and the miRNAs. (b) WT and MyoD^−/− myoblasts were subjected to adipogenic induction and differentiation as shown in the schematics. (c) Representative images of ORO staining of adipocytes. (d and e) qPCR analysis of adipogenic markers (d) and miRNAs (e). Passage 3 primary myoblasts (purity > 95%) from MyoD^−/− and WT mice were used. Error bars represent mean with SD of 3–5 independent biological replicates.^⁎P < 0.05, ^⁎⁎P < 0.01 (Student's t-test).