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. 2020 Sep 23;7(20):2000681. doi: 10.1002/advs.202000681

Figure 7.

Figure 7

PKB/Akt signal activates CSN6 via phosphorylation to regulate FOXO4‐mediated gene expression of SGOC network. A) HCT116 cells were treated with different concentrations of MK2206 and cell viability was measured by CCK8. Values represent average ± s.d., n = 8. B) Real‐time quantitative PCR analysis of Glut1 in indicated cell lines was performed after the treatment of MK2206. Bars represent average ± s.d., n = 3, one‐way ANOVA, *P < 0.05. C) 293T cells were transfected with the indicated CSN6 and CSN6 S60A expression vectors. Cell lysates were immunoblotted with indicated antibodies. D,E) 293T cells were transfected with the indicated expression vectors. Cells were treated with MG132 for 6hr before harvesting, and lysed in denaturing buffer (6 m guanidine–HCl). The cell lysates were then pull down (PD) with nickel beads and immunoblotted with indicated antibodies. F) Real‐time quantitative PCR analysis of serine pathway genes (top) and Glut1 gene (bottom) in HCT116 transfected with indicated expressing plasmids. Bars represent average ± s.d., n = 3, one‐way ANOVA, *P < 0.05. G) Real‐time quantitative PCR analysis was performed to measure mRNA levels of SGOC pathway genes in HCT116 (left) and DLD1 (right) cells treated with MK2206. Bars represent average ± s.d., n = 3, one‐way ANOVA, *P < 0.05.