miR-30a/b/c promote differentiation of myoblasts <i>in vitro</i>.

<p><b>(A)</b> Quantification of MYOG+ nuclei (n ≥ 9 images) in C₂C₁₂ myoblasts transfected with the indicated pre-miRs at 10nM final concentration. 24 hours after transfection, cells were stained with 4’-6-diamidino-2-phenylindole (DAPI) and anti-MYOG antibodies (red).**<i>P</i>≤0.001 compared to pre-miR-control cells, error bars = SEM. Below, representative images of fluorescence and brightfield microscopy. Scale bars = 200μm. <b>(B)</b> Quantification of % MyHC+ area (n ≥ 9 images) from C₂C₁₂ cells transfected with the indicated antimiRs at 10nM final concentrations. 48 hours following withdrawal of serum from the medium, cells were stained with anti-myosin heavy chain (MyHC) antibodies (green) and DAPI. *<i>P</i>≤0.05 compared to antimiR-control, error bars = SEM. Below, representative fluorescence images. Scale bars = 400μm. <b>(C)</b> Quantification of proliferating cells using 5-ethynyl-2’-deoxyuridine (EdU) analysis. 24 hours after electroporation of pre-miR-30a-5p and scrambled controls at indicated concentrations, C₂C₁₂ cells were pulsed with EdU for 2 hours, then stained for EdU incorporation and DAPI. *<i>P</i>≤0.05 vs. scrambled control at equivalent concentrations. Error bars = SEM Right, representative fluorescent images, scale bars = 200μm.