Fig. 4
From: C3G, through its GEF activity, induces megakaryocytic differentiation and proplatelet formation
GATA-1 regulates C3G expression in Dami cells. a Dami cells with a permanent GATA-1 knockdown (shGATA-1) or overexpression of the fused protein V5-GATA-1 (a gift from Dr. Laura Gutiérrez, University of Oviedo), as well as their control cells (expressing empty vectors) were used. Representative flow cytometry plots of the expression of CD41 marker in Dami cells treated with 100 ng/ml PMA for 96 h. Histograms represent the mean ± SEM of the fluorescence intensity (relative units) of CD41 from 3 to 5 independent experiments of each clone. b Histograms represent the mean ± SEM of the percentage of polyploidy (>4n) Dami cells stimulated with PMA for 96 h (n = 5). c Ectopic expression of GATA-1 in Dami cells induces C3G expression. Left panel: Western blot showing the expression of V5-GATA-1 protein or endogenous GATA-1 in Dami cells. V5-GATA-1/β-actin or endogenous (End.) GATA-1/β-actin ratios are shown. Values were normalized against endogenous GATA-1 levels in cells transfected with the empty vector. Right panel: C3G expression in Dami cells transfected with V5-GATA-1 or its control, stimulated with 100 ng/ml PMA for 96 h. Normalized C3G/β-actin ratios are shown. d Silencing of GATA-1 abolished PMA-induced C3G expression in Dami cells. Left panel: Western blot showing GATA-1 levels in Dami cells. Values were normalized using β-actin. Righ panel: C3G expression in Dami cells with silenced GATA-1 expression and treated with 100 ng/ml PMA for 96 h. Normalized C3G/β-actin ratios are shown