Treatment with MI-773 causes changes in p53 status.

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Treatment with MI-773 causes changes in p53 status. Treatment with MI-773 causes changes in p53 status. A, Western blot analysis showing basal levels of proteins p53, MDM2, Bcl-xL, and Bcl-2 in UM-HACC-2A, -2B, -5, and -6 cells. B, Western blots for p53, MDM2, p21, PUMA, BAX, and Bcl-2 protein expression in UM-HACC cells exposed to predetermined IC50 values of MI-773, cisplatin, or vehicle control for 72 hours. C, Representative photomicrographs of UM-PDX-HACC-5 histologic sections stained for p53 (brown) and counterstained with hematoxylin (200×). Nuclear p53 is observed in mice treated with vehicle control or cisplatin, while both nuclear (NE) and cytoplasmic (CE) expression of p53 is evidenced in animals exposed to MI-773 as a single agent or combined with cisplatin (details). D, Western blot analysis for p53 and MDM2 expression in UM-HACC-5 cells exposed to MI-773 (0.1 μmol/L) and/or cisplatin (0.2 μmol/L) and submitted to a subcellular fractionation assay, to evaluate the nucleus and cytoplasm compartments in separate. E, Representative photomicrographs obtained by immunofluorescence for p53 (green), which is located in the cytoplasm (detail) of UM-PDX-HACC-5 cells treated with MI-773 as single drug or combined with cisplatin (400×). Scale bar, 50 μm. Felipe Nör et al. Clin Cancer Res 2017;23:1036-1048 ©2017 by American Association for Cancer Research