Fig. 5: Influence of TCP1 knockdown on the stability of c-Myc and AKT/GSK-3β and ERK signalling pathway components.

a–c TCP1 was knocked down, and the levels of c-Myc, p-c-MycT58, p-c-MycS62, TCP1, AKT, p-AKTS473, GSK-3β, and p-GSK-3βS9 in Huh-7 and SMMC-7721 cells were detected by WB (a). The same detection markers were assessed in PANC-1 and BxPC-3 cells (b). The phosphorylation levels of AKTS473, GSK-3βS9, and ERK, especially the p-AKTS473/AKT, p-GSK-3βS9/GSK-3β and p-ERK/ERK ratios, were significantly reduced with TCP1 knockdown but increased with TCP1 overexpression (c). d, h The GSK-3β inhibitor AR-A014418 was used to confirm the effect of TCP1 on the AKT/GSK-3β axis. Huh-7 cells were treated with 200 nM and 15 μM AR-A014418, and SMMC-7721, PANC-1, and BxPC-3 cells were treated with 200 nM and 5 μM AR-A014418; then, the levels of p-GSK-3β Y216 and c-Myc were detected. e, g, i AR-A014418 was used in scramble and shTCP1 cells to measure c-Myc protein levels. f, j AKT inhibitor VIII (AKT iVIII) was used to determine the relationship between TCP1 and the AKT/GSK-3β pathway. Huh-7 cells were treated with 1 and 10 μM AKT iVIII, SMMC-7721 cells were treated with 1 and 5 μM AKT iVIII, and PANC-1 and BxPC-3 cells were treated with 5 and 10 μM AKT iVIII, and the expression levels of TCP1, p-AKTS473 and c-Myc were detected. The data are shown as the means ± SD from three independent experiments. *P < 0.05, **P < 0.01, between the indicated groups.