Fig. 4: SKP2 suppression leads to myogenic differentiation.

a Representative western blot on cells transfected with either Scrambled (siSCR) or SKP2 siRNA (siSKP2). Vinculin, GAPDH and β-Actin are the loading controls. b mRNA levels of MYOG, MYOD1 and CDKN1C on cells treated as in (a) were reported as fold increase over siSCR (1 arbitrary unit, not reported). Data presented as mean values ± SD, Student’s two-tailed t-test. c mRNA levels of MyH2 on cells treated as in (a) in growth medium (GM) for 4 days post-transfection were expressed as fold increase over siSCR (1 arbitrary unit). Data presented as mean values ± SD, Student’s two-tailed t-test. d Representative immunofluorescence on cells treated as in (a) for 6 days in GM showing expression of Myosin Heavy Chain (MyHC) (green). DAPI (blue) stained nuclei. Scale Bar = 100 μm. e Histogram depicts the quantification of MyHC-positive cells treated as in (d). Data presented as mean values ± SD, Student’s two-tailed t-test. f Representative western blot on cells infected with either shSCR, shSKP2.1 or shSKP2.2. Vinculin is the loading control. g Representative immunofluorescence on cells treated as in (f) for 6 days, showing expression of MyHC (green). DAPI (blue) stained nuclei. Scale Bar = 100 μm. h Histogram depicts the quantification of MyHC-positive cells treated as in (g). Data presented as mean values ± SD, two-way ANOVA. i Representative western blot on cells treated with SMIP004 20 μM for 168 h. Vinculin is the loading control. j mRNA levels of MYOG, MYOD1 and CDKN1C on cells treated as in (i) were reported as fold increase over DMSO (1 arbitrary unit, not reported). Data presented as mean values ± SD, Student’s two-tailed t-test. k mRNA levels of MyH2 on cells treated as in (i), cultured in GM, were expressed as fold increase over DMSO (1 arbitrary unit). Data presented as mean values ± SD, Student’s two-tailed t-test. l Representative immunofluorescence of cells treated as in (i) showing expression of MyHC (green). DAPI (blue) stained nuclei. Scale Bar = 100 μm. m Histogram depicts the quantification of MyHC-positive cells treated as in (i). Data presented as mean values ± SD, Student’s two-tailed t-test. All presented data derive from n = 3 independent experiments. Source data are provided as a Source Data file.