Figure 5.

SNX16 activates the c‐Myc signaling pathway by inhibiting eEF1A2 degradation in CRC. (A) The expression of SNX16 and eEF1A2 in SNX16‐knockdown or SNX16‐overexpressing cells was measured by western blotting. Tubulin was used as the loading control. (B) The expression of SNX16 and eEF1A2 in SNX16‐knockdown or SNX16‐overexpressing cells was measured by qRT‐PCR. GAPDH was used as the loading control. (C) eEF1A2 levels were determined in SNX16‐knockdown HT29 cells and SNX16‐overexpressing SW480 cells before and after MG132‐mediated stimulation by western blotting. (D) SNX16‐knockdown HT29 cells and SNX16‐overexpressing SW480 cells were exposed to CHX (20 μg·mL⁻¹) at the indicated time point, and degradation of eEF1A2 was detected by western blot analysis. (E) SNX16‐knockdown HT29 cells and SNX16‐overexpressing SW480 cells were treated with MG132, and the level of ubiquitin‐bound eEF1A2 was then measured. (F) Upregulation of c‐Myc induced by SNX16 was attenuated upon knockdown of eEF1A2 in SNX16‐overexpressing cells. (G) MTT assay. The results are shown as the means ± SEMs (n = 5), ****P < 0.0001. (H) Colony formation assays. Results are shown as mean ± SEM (n = 3), *P < 0.05, ***P < 0.001