MiR-301a promotes embryonic stem cell differentiation to cardiomyocytes

Figure 3 MiR-301a promotes mouse embryonic stem cell differentiation to cardiomyocytes. A: Quantitative real-time PCR analysis of the gene expression for mid-stage cardiac markers, including GATA-4, TBX5, and MEF2C, and late-stage cardiac markers, including α-actinin, sarcomeric MHC, and MLC, in cells at day 4 (embryoid body formation), day 8 (cardiac differentiation), and day 12 (immature cardiomyocytes) during mouse embryonic stem cell differentiation as indicated in Figure 2A. The gene expression levels are shown as the fold change compared to control cells at day 4. Data are presented as the mean ± SEM (n = 3); B: Western blot analysis of the cardiac markers, including GATA-4, MEF2C, NKX2.5, CTNT, and α-actinin, in cells at day 12 after mouse embryonic stem cell differentiation with or without overexpression of miR-301a. A semiquantitative analysis (cardiac markers normalized on GAPDH, miR-301a group vs control group) was applied and shown; C: Quantitative analysis of the percentage of beating embryoid bodies at different time points demonstrated earlier initiation and more embryoid body beating in the miR-301a group than in the control group. All of the embryoid bodies in three independent dishes in each group (141 embryoid bodies in the control group and 131 embryoid bodies in the miR-301a group) were calculated at each time point. Data are presented as the mean ± SE (n = 3). ^aP < 0.05, ^bP < 0.01.