Fig. 1: FASN is a specific mutp53-binding protein.
From: Lipogenic enzyme FASN promotes mutant p53 accumulation and gain-of-function through palmitoylation
a Ectopic FASN protein specifically interacted with ectopic hotspot mutp53 but not wtp53 in cells. p53-/- HCT116 cells co-expressing FASN-Flag and ectopic wtp53 or different hotspot mutp53 were employed for co-IP assays. -: control vectors. WT: wild-type. b Co-IP analysis of the interaction of endogenous FASN with different endogenous hotspot mutp53 in SK-BR3, MDA-MB468, and LS1034 cells. c Endogenous FASN protein specifically interacted with endogenous hotspot mutp53 proteins, but not wtp53, in cells. Isogenic HCT116 cells and MEFs with different status of p53 were used for co-IP assays. d The DBD domain of R175H mutp53 is required for mutp53-FASN interaction. Left: schematic representation of vectors expressing full-length (FL) and serial deletion mutants of R175H HA-mutp53. AD: transactivation domain; DBD: DNA-binding domain; TD: tetramerization domain; CTD: C-terminal domain. Right: p53-/- HCT116 cells co-expressing FASN-Flag and FL or deletion mutants of R175H HA-mutp53 were used for co-IP assays. e The direct interaction between mutp53 and FASN proteins analyzed by in vitro His pull-down assays using purified recombinant FASN-Flag and His-mutp53 (R175H) or His-wtp53 proteins. f The in situ interaction between endogenous mutp53 and FASN proteins in cells analyzed by the proximity ligation assay (PLA). Left panels: Represented PLA images. Red: PLA signals of the mutp53-FASN interaction in cells. Scale bar: 10 μm. Upper right panel: mutp53 KO by CRISPR/Cas9 in SK-BR3 cells. Lower right panel: the quantification of PLA signals in the nuclear and cytoplasmic compartments. Totally 200 cells were counted in 8 different areas. In a–f, all Western blot data represent three repeats with similar results. Source data are provided as a Source Data file.
