Colo858

BRAF-mutated melanoma cell lines used in this study were obtained from the Massachusetts General Hospital Cancer Center with the following primary sources: COLO858 (ECACC) and MMACSF (Riken Bioresource Center). Each cell line was independently authenticated by Short Tandem Repeats (STR) profiling by ATCC. COLO858 cells were grown in RMPI 1640 (Corning cellgro, Cat. 10–040 CV), and MMACSF cells were grown in DMEM/F-12 (Thermo Fisher Scientific, Cat. 11330–032). For both cell lines, growth media were supplemented with 5% fetal bovine serum (Thermo Fisher Scientific, Cat. 26140–079) and 1% sodium pyruvate (Thermo Fisher Scientific, Cat. 11360–070). We added penicillin and streptomycin at 100 U/ml (Thermo Fisher Scientific, Cat. 15140–122) and plasmocin at 0.5 μg/ml (InvivoGen, Cat. ant-mpp) to all growth media. Cells were engineered to stably express H2B-Venus and mCherry-Geminin fluorescent reporters as described previously [21 (link)]. Engineered and parental cell lines were confirmed to grow at comparable rates in the absence of any treatment or in the presence of different concentrations of BRAF inhibitor Vemurafenib over 72 hours of treatment.

Melanoma cell lines used in this study were obtained from the Massachusetts General Hospital Cancer Center with the following primary sources: COLO858 (ECACC), A375, C32, WM115, SKMEL28, and WM1552C (ATCC), LOXIMV1 (DCTD Tumor Repository, National Cancer Institute), MMACSF (RIKEN BioResource Center), and MZ7MEL (Johannes Gutenberg University Mainz). C32, MMACSF, SKMEL28, and WM115 cell lines were grown in DMEM/F12 (Invitrogen) supplemented with 5% fetal bovine serum (FBS) and 1% sodium pyruvate (Invitrogen). COLO858, LOXIMVI, MZ7MEL, and WM1552C cell lines were grown in RMPI 1640 (Corning cellgro) supplemented with 5% FBS and 1% sodium pyruvate (Invitrogen). A375 cells were grown in DMEM with 4.5 g/l glucose, l‐glutamine, and sodium pyruvate (Corning cellgro), supplemented with 5% FBS. We added penicillin (50 U/ml) and streptomycin (50 μg/ml) to all growth media.