Figure 5. Oncogenic B-Raf promotes melanosphere formation.
(A) Melanosphere formation was assessed for Hs936T cells, Hs936T cells stably expressing either an empty pBABE vector (EV) or pBABE vector containing oncogenic B-RafV600E, and Hs936T cells stably expressing pBABE-B-RafV600E and treated with 5 μM PLX-4032. (B) Following the 10-day melanosphere assay, cells were collected and melanospheres dissociated by incubation with TrypLE and counted. Error bars represent the SEM of four biological replicates of each melanosphere assay. (C) Cells were grown in adherent conditions and protein levels for Nrf2 and B-Raf were assessed by immunoblot analysis. Band intensities were measured and normalized to tubulin. Error bars represent the SEM of the three biological replicates used for immunoblot analysis. Statistical significance was determined using the standard Student’s t-test. *P < 0.05, ***P < 0.001.