FIGURE 7.

Effect of 124mut on TRAF6 ubiquitination, pNF‐κB, and VEGF‐C expression. (A) Schematic diagram of ubiquitin. (K: lysine; R: arginine). (B, C) To detect the effect of the 124th ubiquitination site mutation on TRAF6 ubiquitination, TRAF6 or TRAF6 124mut construct were transfected into colorectal cancer (CRC) cells with various ubiquitin plasmids, respectively. Ubiquitinated TRAF6 was detected with the anti‐HA antibody from immunoprecipitated TRAF6 protein. (D, E) Western blot analysis of pNF‐κB and VEGF‐C protein levels in SW480 cells after transfection with pCDH‐3×Flag‐TRAF6 or pCDH‐3×Flag‐TRAF6 124mut plasmids. GAPDH served as a loading control. The relative protein expression levels of TRAF6, pNF‐κB, and VEGF‐C in SW480 and HCT116 cells were quantified. Data are shown as mean ± SD. *P < 0.05; **P < 0.01