Fig. 2

p53R248Q and p53R273C mutants positively regulates the expression of both HER2 mRNA and protein. a-b The mutant p53R248Q harbored in the OVCAR-3 cell line was inhibited by specific shRNAs for p53 (shp53), as indicated in Materials and Methods. After 48 h, transfected OVCAR-3 cells were processed to determine a HER2 mRNA relative levels by RT-qPCR and b HER2 protein levels by Western Blot. c-d The Saos-2 cell line (p53-null) was stably transfected with mutant p53R248Q, or with the empty vector, as indicated in Materials and Methods. After stable selection, c HER2 mRNA expression assay was performed by RT-qPCR, and d protein expression was measured by Western Blot. The RT-qPCR results are the means of at least three independent experiments ± SEM and the image of the blot is representative of three independent experiments. Statistical analysis was performed with t-student test by comparing the results obtained in: shp53 vs scramble transfected OVCAR-3 cells; or p53R248Q vs empty vector transfected Saos-2 cells: * p < 0.05. e-f The expression of p53R273C mutant in C33A cell line was transiently inhibited by short-hairpin RNA against p53 (shp53), under the conditions indicated in Materials and Methods. After 48 h the cells were processed for: e HER2 mRNA expression assay by RT-qPCR, and f protein expression assay by Western Blot. g-h The Saos-2 cell line (null-p53) was stably transfected with mutant p53R273C, or with empty vector, as indicated in Materials and Methods. Transfected cells were processed after stable selection for: g HER2 mRNA expression assay by RT-qPCR, and h protein expression assay by Western Blot. The RT-qPCR results are the means of at least three independent experiments ± SEM and the image of the blot is representative of three independent experiments. Statistical analysis was performed with t-student test by comparing the results obtained in: shp53 vs scramble transfected C33A cells; or p53R273C vs empty vector transfected Saos-2 cells: * p < 0.05