Boysen et al. Supplemental Figure 4
Boysen et al. "Hsp70 and Hsp90-mediated Regulation of the Conformation of p53 DNA binding domain and p53 Cancer Variants", Supplemental Figure S4 related to figure 2: p53-DBD wild type and mutant proteins remain soluble when pre-incubated at 37°C in the presence of Hsp70 and Hdj1. In the absence of chaperones (left panel) or in the presence of Hsp70, Hdj1 and ATP (right panel) p53-DBD-wt was pre-incubated for 70 min at 37°C, p53-DBD-R175H 10 min at 37°C, p53-DBD-R248Q and p53-DBD-R249S for 30 min at 37°C. Subsequently, aggregated and soluble (S) fractions were separated by centrifugation and supernatant (S) and pellet (P) were analyzed by SDS-polyacrylamid gel electrophoresis and Coomassie Brillant Blue staining.