Figure 4.

TRIM21 is the E3 ligase for GAC. A, B Indicated plasmids were transfected into H1299 cells. Interaction between GAC and TRIM21 were detected by immunoprecipitation and western blot assay. C-E Indicated plasmids were transfected into H1299 cells. The levels of GAC ubiquitination were detected by immunoprecipitation and western blot assay. F-H Indicated plasmids and siRNAs were transfected into H1299 cells. The levels of GAC ubiquitination were detected by immunoprecipitation and western blot assay. WCL: whole cell lysate. I Indicated siRNAs were transfected into H1299 cells. The protein expression was determined by western blot and glutaminase activity assay was performed. J Indicated plasmids were transfected into H1299 cells. The protein expression were determined by western blot and glutaminase activity assay was performed. Data are showed as mean ± SD, n=3. **P < 0.01.