Figure 2.

Characterization of proliferation behavior. A-C: Proliferation rates of cells as assessed by manual counting. D-E: Staining of senescence-associated β-galactosidase activity and quantification of senescent cells revealed no relevant amounts of cellular senescence to be present in RBW-1 or other clones. Microscopy scale: 150 μm × 100 μm. F: In 10% FBS all cell lines showed similar cell cycle patterns in flow cytometry. Reduction of serum led to aberrant patterns for RBW-1 compared to BRAF-mutant clones. G: Quantification of sub G1 fraction revealed high amounts of cell death in RBW-1 after incubation with 0.5% FBS. H: Apoptosis was analyzed by Western blotting for cleaved caspase 3, p53, and PUMA.