Hs294T (p53WT), SK-Mel-2 (p53 mutant), SK-Mel-5 (p53WT) and SK-Mel-28 (p53 mutant) cells were treated with 1 µM MLN8237 for 6 or 12 days. After treatment, viable cells were counted and compared to the initial cell number. Data indicate mean values ± SD (n = 3) from one representative of three independent experiments. p-value shown represents difference between treated group and day 0 control group (Student's t-test).
Cultures of Hs294T, SK-Mel-2, SK-Mel-5 and SK-Mel-28 cells were treated with MLN8237 or vehicle for indicated time points and apoptosis was analysed by FACS analysis for propidium iodide (PI) and Annexin V staining. Data indicate mean values ± SD (n = 3) from triplicate experiments.
Hs294T cells were treated with 1 µM MLN8237 or vehicle for 5 days and the change in morphology was captured with an AxioVision microscope.
Hs294T cells were treated with 1 µM MLN8237 for 5 days, and senescence was determined by β-galactosidase staining.
Melanoma cell lines with wild-type p53 (Hs294T and SK-Mel-5) or mutated p53 (SK-Mel-2 and SK-Mel-28) were treated with 1 µM or 5 µM MLN8237 for 5 days. After treatment, p53, p63, p73, p21, p16 and GAPDH were analysed by Western blot.
Hs294T and SK-Mel-28 cells were treated with 1 µM MLN8237 or vehicle in the presence or absence of the p53 inhibitor pifithrin-α (PTF-α, 5 µM) or DMSO for 5 days. After treatment, β-galactosidase staining was performed. All experiments were conducted at least three times independently with reproducible results.
