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. 2023 Feb 7;120(7):e2212940120. doi: 10.1073/pnas.2212940120

Fig. 3.

Fig. 3.

Altered biochemical properties of hypomorphic p53 proteins. (A) WT, P47S, and Y107H LCLs were treated with or without 1 mM BMH for 30 min followed by immunoblotting for p53. (B) WT, P47S, and Y107H LCLs were treated with vehicle or 1 μg/mL ATO for 16 h and then fixed with or without 1 mM BMH for 30 min followed by immunoblotting for p53. (C) WT or Y107H LCLs were untreated or treated with 10 μM Nutlin-3a (Nut), 1 μM doxorubicin (Dox), or 1 μg/mL ATO for 24 h. Whole-cell lysates were analyzed by western blotting with the indicated antibodies. (D) Cytosolic and nuclear fractions isolated from WT, P47S, and Y107H LCLs were analyzed by western blotting with the indicated antibodies. Histone H3 and GAPDH served as nuclear and cytosolic controls, respectively, to confirm efficiency of subcellular fractionation. (E and F) Chromatin-bound proteins were extracted from nuclear pellets isolated from WT, (E) P47S, and (F) Y107H LCLs using sequentially increasing salt concentrations (0 to 500 mM NaCl) and analyzed by western blotting for p53.