Fig. 4
Expression and subcellular localization of MKL1 and SRF protein in ARMS cells. a Confocal images of MKL1 and SRF expression and subcellular localization in RH30 cells with or without overexpressed PAX3-FOXO1 protein. Protein expression of MKL1 (green) and SRF (red) and the colocalization or association (yellow) between them were attenuated with ectopic PAX3-FOXO1 expression, but almost not changed for their subcellular distribution by immunofluorescence staining. b Analysis of total MKL1 and SRF expression in RH4 cells with increasing amount of PAX3-FOXO1 protein expression. Left: MKL1 and SRF levels determined by western blot. Right: the relative expression of MKL1 and SRF protein shown on the left. c Analysis of total MKL1 and SRF expression in RH4 cells transfected by siRNAs against PAX3-FOXO1 gene. Left: Expression of MKL1 and SRF determined by western blot. Right: the relative expression of MKL1 and SRF protein shown on the left. d Distribution of SRF and MKL1 proteins in cytoplasmic fraction of RH30 cells transfected with increasing amount of PAX3-FOXO1 plasmid. Left: Expression of MKL1, SRF and PAX3-FOXO1 protein determined by western blot analysis. Right: the relative quantification analysis of MKL1, SRF and PAX3-FOXO1 protein shown on the left. α-Tubulin served as a loading control. e Distribution of SRF and MKL1 expression in nuclear fraction of RH30 cells transfected with increasing amount of PAX3-FOXO1 plasmid. Left: Expression of MKL1, SRF and PAX3-FOXO1 protein determined by western blot. Right: the relative quantification analysis of MKL1, SRF and PAX3-FOXO1 protein shown on the left. TBP served as a loading control for nuclear fraction. f Analysis of total RhoA expression in RH4 cells with a different dose of PAX3-FOXO1 plasmid transfected. Left: RhoA level determined by western blot. Right: the relative expression of RhoA protein shown on the left. g Analysis of total RhoA expression in RH4 cells transfected by siRNAs against PAX3-FOXO1 gene. Left: Expression of RhoA determined by western blot. Right: the relative expression of RhoA protein shown on the left