Figure 2. Cisplatin induces prohibitin accumulation and cell death in different melanoma cells.
Cells were treated with 25 μM of cisplatin for 24 h. (A) Cisplatin treatment induced increase in the frequency of hypodiploid cells in Mel 85, LB373, MeWO, MZ2Mel and SKMel 37 cell lines. Cell death was also determined using both JC-1 staining and annexin V/PI double staining for LB373 cell line (Supplementary Figure 2). Protein extracts of LB373 (B), Mel85 and SKMel 37 (C) cells, either treated (+) or not (−) with cisplatin were separated in SDS-PAGE and blotted for analysis of PHB and beta-actin accumulation. Protein accumulation was measured by densitometry using ImageJ software. The ratio of PHB/beta-actin accumulation was fixed as 1 for all the control conditions (non-treated cells). Values > 1 indicate relative accumulation of PHB after cisplatin treatment. Representative experiment of three independent assays.