Figure 2
PEITC induces a ‘WT-like’ conformational change in p53R175 mutant protein. (a) ELISA to determine the effect of PEITC on conformation of recombinant-purified GST-p53R175H by using conformation-specific antibodies PAB240 (mutant-specific) and PAB1620 (WT-specific). (b) SK-BR-3 cells were treated with DMSO or 4 μM PEITC for 6 h. Immunofluorescence of the cells was performed using PAB240 and PAB1620 antibodies. The A549 cell line used as a control showed that p53 WT conformation was not changed by PEITC. The H1299 cell line was used as a control for anti-p53 antibodies. All scale bars represents a size of 20 μm. (c) Quantification of PAB240 and PAB1620 staining shown in panel (b). ***P≤0.0001 for PAB240 and PAB1620. (d) Immunoprecipitation of the p53 mutant protein from SK-BR-3 cell lysates using PAB240 antibody and detected by p53 (FL393) antibody
