C3G is essential for the maintenance of the pluripotency factor network. a Volcano plot showing changes in the global expression of genes in C3G knockout cells compared to WT cells. Dotted lines indicate the significance of p < 0.05. Significantly altered expression of genes involved in pluripotency, differentiation, and adhesion are highlighted. b qPCR analysis of indicated pluripotency factors in vector control (VC) and C3G KO cells relative to WT cells.*P < 0.05, **P < 0.01, ***P < 0.001; n = 3 c Western blots showing expression of the indicated proteins in WT, VC, and C3G KO clones. Quantification averaged from three independent experiments relative to levels seen in WT is represented in the bar diagram. ACTIN was used as a loading control. d Western blot showing pSTAT3, and pERK levels. Quantification of levels normalized relative to respective total proteins is shown in the bar diagram, p < 0.001; n = 3. e Quantification of differential expression of indicated differentiation factors determined by qPCR. *P < 0.05, **P < 0.01, ***P < 0.001, n = 3. f Colony morphology of WT, KO clone D1, and D1 clone upon transient expression of C3G (R-24). g Western blots showing expression levels of indicated proteins in KO clone (D1) upon the rescue of C3G expression. Quantification relative to expression in WT cells is shown in the bar diagram. *P < 0.05, **P < 0.01 ***P < 0.001; n = 3.