Extended Data Fig. 2: ME2 controls mutant p53 stability.
From: Malic enzyme 2 maintains protein stability of mutant p53 through 2-hydroxyglutarate
a, mRNA levels of p53R280K in MDA-MB-231 cells treated with Ctrl or ME2 siRNA (siME2) for 48 hours (n = 3 biological independent samples). b, and d, Sequencing analysis of mutations in TP53 gene in MDA-MB-231 cells (b) and U251MG cells (d). Blue shadings indicate the mutation positions. An AGA-to-AAA mutation in TP53 gene has been detected in MDA-MB-231 cells, and a CGT-to-CAT in TP53 gene in U251MG cells. c, Western blot analysis of p53 expression in MDA-MB-231 cells transfected with Ctrl, ME1, or ME2 siRNA for 48 hours. e, U251MG cells transfected with Ctrl or ME2 siRNA for 48 hours were treated with 200 µg/mL CHX for another indicated time points in the presence or absence of 10 µM MG132. Protein expression was analyzed by western blot (n = 3 biological replicates). Relative p53/actin ratios are shown. f, U251MG cells transfected with Ctrl, ME2 and/or Mdm2 siRNA as indicated for 48 hours were treated with 200 µg/mL CHX for another indicated time points. Protein expression was analyzed by western blot. Relative p53/actin ratios are shown. Data in a, e and f are mean ± SD, P values were determined by unpaired two-tailed Student’s t-tests.
