Fig. 1
From: p53-mediated adaptation to serine starvation is retained by a common tumour-derived mutant
The R248W mutant retains wild-type ability to engage p21 and MDM2 and promotes adaptation to serine and glycine (SG) deprivation. a Western blot for p53 in HCT116 cells with parental WT p53 (WT), CRISPR-mediated p53 deletion (KO), or p53 KO cells reconstituted with mutant p53 expression constructs R175H, R248W, or R273H (175, 248, 273). HSP90 expression used as a loading control. b p53 protein in wild-type (IP by pAB1620) and mutant/unfolded (IP by pAb240) conformation in the indicated cell lines, detected by immunoprecipitation followed by Western blot. c Western blot for p53, MDM2, and p21 in the indicated cell lines treated with the MDM2 inhibitor RG7388 (+) (5 μM) or DMSO vehicle control (−) for 24 h. d The indicated cell lines were cultured in full medium, full medium containing the MDM2 inhibitor nutlin-3A (10 μM), or SG-depleted medium for 8 days. e Cell counts of the indicated cell lines cultured in full medium, full medium containing the MDM2 inhibitor nutlin-3A (10 μM), or SG-depleted medium for 8 days. Data are represented as the mean of triplicate wells ± SEM for each condition. f Western blot for MDM2 and p21 in the indicated cell lines cultured in full medium (day 0) before being switched to grow in SG-depleted medium for 1 or 2 days. Note: this blot was re-probed for PSAT-1 and PSPH and shown in Fig. 2c. The ACTIN loading control blot is therefore the same in both figures