Extended Data Fig. 4: C269 palmitoylation of mutant IDH1 was not mediated by ZDHHC family enzymes.
From: Autopalmitoylation of IDH1-R132H regulates its neomorphic activity in cancer cells
a-c, Screening of mouse Zdhhc-PATs that are responsible for IDH1mt palmitoylation. HEK293A cells were co-transfected with Flag-IDH1mt and HA-Zdhhcs. Palmitoylation signals of IDH1mt were determined by streptavidin pulldown and western blot analysis. d, Analysis of palmitoylation levels of IDH1mt 3CS (C73S/C114S/C379S) after knocking down endogenous ZDHHC14 or 23 using shRNA. Bar graph showing knockdown efficiency of ZDHHC14 or ZDHHC23 at the mRNA level (n = 3 independent experiments). Data were shown as mean ± SEM. Statistical significance was assessed using an unpaired, two-sided Student’s t-test. e-f, Analysis of palmitoylation levels of IDH1mt 3CS after knocking down endogenous ZDHHC7 (e) or 9 (f) using siRNAs. g, Analysis of palmitoylation levels of IDH1mt 3CS after individually overexpressing Zdhhc7/9/14/23. h, Co-IP assays showing there were no interactions between IDH1mt 3CS and Zdhhc7, 9, 14 or 23. The experiments in a-c and e-h were repeated independently three times with similar results, and representative results are presented.
