Fig. 6.

(a) Mutational status of NRAS, p53, and BRAF genes in melanoma cell lines was tested using qRT-PCR analysis. Cell lines WC00046, WC00060, and WC00081 are represented by solid black, solid white, dotted black bars. (b)Western blot analysis of Bcl-2 (1 : 1000), PARP 1 (1 : 2000), and caspase-3 (1 : 2000) was carried out on 12% SDS-PAGE gels and the whole-cell extracts were transferred onto a PVDF membrane. The membranes were washed and incubated with a 1 : 5000 dilution of horseradish peroxidase-conjugated donkey anti-mouse IgG. The blots were washed and developed using the ECL system. (c) Analysis of cyclin D1 and cyclin D2 expressions in melanoma cell lines was carried out with PE-labeled antibodies using flow cytometry. A total of 1×10⁶ cells from control and irradiated groups were harvested from T-25 flasks 24 h after radiation by centrifugation at 1200 rpm for 7 min, and cell pellets were washed with ice cold PBS. The Per Fix-nc kit was used for fixing, permeabilizing, and preparing the cells for the assay. HEM, human epidermal melanocytes; PVDF, polyvinylidene difluoride; qRT-PCR, quantitative real-time reverse transcriptase PCR.