The p53-Stabilizing Compound CP-31398 Enhances Ultraviolet-B-Induced Apoptosis in a Human Melanoma Cell Line MMRU Yvonne Luu, Gang Li, Dr Journal of.

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The p53-Stabilizing Compound CP-31398 Enhances Ultraviolet-B-Induced Apoptosis in a Human Melanoma Cell Line MMRU Yvonne Luu, Gang Li, Dr Journal of Investigative Dermatology Volume 119, Issue 5, Pages 1207-1209 (November 2002) DOI: 10.1046/j.1523-1747.2002.19517.x Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 CP-31398 enhances UVB-induced apoptosis by increasing p53 protein levels in MMRU cells. MMRU cells were irradiated with 0, 40, 60, or 80 mJ per cm2 of UVB followed by treatment with 0 or 8 μg per ml of CP-31398 for 24 h. (A) Western analysis of p53 level with DO-1 antibody (Santa Cruz Biotechnology, Santa Cruz, CA) and β-actin (BD Pharmingen, Canada) as loading control. The fold of p53 induction was determined by densitometry. (B) Cells were stained with propidium iodide and the pre-G1 population was determined by flow cytometry. (C) DNA was extracted and analyzed on a 2% agarose gel. Journal of Investigative Dermatology 2002 119, 1207-1209DOI: (10.1046/j.1523-1747.2002.19517.x) Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 CP-31398 activates the mitochondrial pathway during UVB-induced apoptosis. MMRU cells were irradiated with 0 or 40 mJ per cm2 of UVB followed by treatment with 0, 8, or 10 μg per ml of CP-31398 for 24 h. (A) Western analysis of Bax and cytochrome c levels with anti-Bax (N-20) (Santa Cruz Biotechnology) and anticytochrome c (BD Pharmingen) antibodies. (B) Cells were stained with MitoCapture™ solution containing a cationic dye and then visualized using fluorescent microscopy. The graph shows quantification of the percentage of green apoptotic cells based on the changes in the mitochondrial membrane potential. At least 500 cells from five random fields were counted. Data represent mean±SD from three independent experiments. Paired two-tailed Student’t test. *Significance: p<0.05, compared with UVB alone. (C) Western analysis of caspase-9 and caspase-3 levels. β-actin was used as loading control. The relative expression levels of procaspase-9 and procaspase-3 were determined by densitometry. Journal of Investigative Dermatology 2002 119, 1207-1209DOI: (10.1046/j.1523-1747.2002.19517.x) Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions