Figure 5
Evaluation of p53 and p21 expression and localization in canine melanoma cell lines after KPT-335 treatment. A. Canine melanoma cell lines were treated with DMSO or 1 μM KPT-335 for 24 hours prior to collection. Protein lysates were generated, separated by SDS-PAGE and Western blotting for p53, p21 and β-actin were performed. B. Canine melanoma cell lines were treated with DMSO or KPT-335 at 0.1 μM or 1 μM for 4 hours (Mel 69) or 24 hours (Mel 23 and Mel 83). RNA was collected and quantitative RT-PCR was performed for MIC-1 and relative expression of MIC-1 was significantly increased in Mel 23 and Mel 69 cells treated with 0.1 μM and 1 μM KPT-335 and MIC-1 was significantly increase in Mel 23 and Mel 69 cells treated with 0.1 μM and 1 μM (p < 0.05). C. Canine melanoma cell lines were treated with DMSO or 1 μM KPT-335 for 24 hours prior to collection. Nuclear and cytoplasmic protein lysates were generated and separated by SDS-PAGE and Western blotting for p53, p21 and β-actin was performed.