FIGURE 7.

Silent information regulator 2 (SIRT2) increased glucose 6‐phosphate dehydrogenase (G6PD) protein stability by reducing G6PD ubiquitination and enhancing G6PD small ubiquitin‐related modifier 1 (SUMO1) modification. A, B, G6PD protein expression levels were analyzed by western blot analysis in stably transfected ACHN and Caki‐1 cells following treatment with 100 μg/mL cycloheximide (CHX) with or without 10 μg/mL MG132 for 0, 6, 9, 12 h. C, Expression levels of G6PD ubiquitination were analyzed by quantitative coimmunoprecipitation (Co‐IP) assays in stably transfected ACHN and Caki‐1 cells. D, Expression levels of G6PD SUMO1 modification were analyzed by quantitative Co‐IP assays in stably transfected ACHN and Caki‐1 cells. E‐G, G6PD activity was measured in ACHN, Caki‐1, and 786‐O cells with DMSO or 2‐D08. H‐J, G6PD dimer and monomer expression level in ACHN, Caki‐1, and 786‐O cells with DMSO or 2‐D08 were analyzed using western blot assays with glutaraldehyde cross‐linking. K, L, Expression levels of G6PD SUMO1 modification and ubiquitination were analyzed by quantitative Co‐IP assays in ACHN, Caki‐1, and 786‐O cells with DMSO or 2‐D08. Data represent three independent experiments, each carried out in triplicate. *P < .05, **P < .01, ***P < .001 vs corresponding controls (unpaired‐sample Student’s t test). IB, immunoblot