Figure 1
From: p63 is required beside p53 for PERP-mediated apoptosis in uveal melanoma
Endogenous PERP expression in UM cells and differential response following TRAIL and UV treatment. Low PERP expression at the (A) transcriptional and (B) protein level in all UM cell lines tested indicated downregulation of PERP in these cells. (A) endogenous PERP mRNA levels determined by qPCR normalised to the level of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) are shown relative to the normalised PERP mRNA level in ARPE19 cells (given an arbitrary value of 1). The mean of three independent experiments along with s.d. is presented. (B) Western blot showing relative protein levels of endogenous PERP in UM cell lysates. A431 cell lysate (human epithelial carcinoma whole-cell lysate; Abcam; ab7909) served as a positive control for the PERP antibody. (C) Differential response of OCM-1 and MEL290 cells to the TRAIL treatment for 36 h at the concentration of 300, 450 and 600 ng ml−1. OCM-1 cells remained viable with normal cell morphology, even at the highest TRAIL concentration. (D) Western blots indicating the level of PERP and DR4 proteins in OCM-1 cell lysates following treatment (36 h) with various concentrations of TRAIL. Non-treated (NT) cells served as control for the TRAIL treatment. (E) Following UV irradiation at the dose of 15 mJ cm−2, OCM-1 cells were collected and lysed at 3, 24 and 48 h post-treatment for immunoblot analysis using PERP antibody. UV-treated Cos-7 cell lysate (Monkey kidney whole-cell lysate; Santa Cruz Biotechnology, Insight Biotechnology Ltd, Middlesex, UK; sc-24666) served as a positive control. GAPDH was used as the loading control in all experiments.
