Anchorage-independent growth of HME16C cell lines. (A) Number of soft agar colonies exceeding 100 μM after 21 days of growth for HME16C cell lines infected with pLRT control, H-RasV12, H-RasV12G37, H-RasV12S35, H-RasV12C40, and Rlf-CAAX-expressing constructs. (B) Representative pictures of colony sizes in soft agar assays. Bar represents 200 μm. (C) Anchorage-independent growth of cell lines grown on ultra-low attachment plates treated with DMSO vehicle (left) or 0.25 μM PD153035 (right) measured using the alamar blue assay. RFU, relative fluorescence units of alamar blue product formed.