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. 2017 Apr 27;6(7):770–779. doi: 10.1016/j.molmet.2017.04.006

Figure 4.

Figure 4

miR-23b and miR-27b regulate the p53 signaling pathway in muscle stem cells. Schematic overview of the proposed interactions between the miR-23b/27b cluster, the p53 pathway and myogenesis (A). P53 protein levels were measured by electrochemiluminescent immunoassay (B–C). Muscle stem cells from T2DM donors were transfected with 50 nM miR-23b or miR-27b mimics, a scrambled, non-targeting oligonucleotide control and for immunoblots also a lipofectamine only control (LF). Cells were harvested 48 h post transfection. Protein levels of MDM2, RCHY1, Cyclin G1 and beta tubulin were determined using immunoblotting. A representative blot is shown (D). mRNA expression of MDM2, RCHY1 and CCNG1 (E) and p21 and RB1 and P53 (F) was measured by qPCR and data was normalized to GAPDH mRNA. Myogenic markers including PAX7 (G), MYOD (H), MYOG (I) and MYH2 (J). Data are means ± SEM. * indicates P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.