Fig. 4
GOLGA7 knockout does not affect NRAS palmitoylation (A) Acyl-RAC assays of HeLa cells ± sgGOLGA7 transfected with GFP-NRASG12D or HeLa cells transfected with GFP-NRASG12D, C181S. Palmitoylation of NRASG12D, C181S mutant is undetectable, while GFP-NRAS.G12D in HeLa cells ± sgGOLGA7 are all shown to be palmitoylated (Palm in IP). The preserved fraction of immunoprecipitants (Neg in IP) show the specificity of the assay. The cleaved fraction (Palm in Supernatant) and preserved fraction of supernatants (Neg in Supernatant) represent non-palmitoylated proteins and the total protein unbound to the resin, respectively. B Statistical quantification of relative palmitoylated protein in (A). Values are the means ± SD from three independent experiments, compared using two-sided Student’s t test. GOLGA7-sg1, ns p = 0.7751; GOLGA7-sg2, ns p = 0.7229; C181S, ***p = 0.0001