Fig. 2: Suppression of SNAI2 activates myogenic differentiation and suppresses stemness in vitro in FN-RMS.

a The level of SNAI2 knockdown by shSNAI2 in RD cells compared to shScr assessed by western blot (Representative blot, n = 3 biologically independent experiments). b Representative images of immunostaining in RD cells stably expressing shScr or c shSNAI2.1 shRNA stained for Myosin Heavy Chain 1 (MyHC, green), MEF2C (red) and DAPI for nuclei (blue). d Quantitation of immunostaining counts as percentage values to total nuclei per image (n = 3 biologically independent experiments, data presented as mean values ± SD, Student’s two-tailed t-test, exact p values are reported in the figure). e qRT-PCR gene expression analysis in RD cells comparing shScr to shSNAI2.1 KD showing early and late myogenic markers (n = 3 biologically independent experiments, data presented as mean values ± SD, Student’s two-tailed t-test, exact p values are reported in the figure). f Representative western blot (n = 3 biologically independent experiments) of muscle differentiation genes in RD shScr vs shSNAI2.1 transient KD cells at 3, 5, 7, and 10 days post puromycin selection. g, h Representative images of sphere formation assays in RD cells containing shScr or shSNAI2.1. i Quantitation of sphere counts in RD cells plated at three densities (10,000, 1000, and 100 per well). n = 3 biologically independent experiments, data presented as mean values ± SD, Student’s two-tailed t-test, exact p values are reported in the figure, Scale Bars, b, g = 100 μM.