Oncogenic Ras^G12V-induced DNA polymerase depletion is associated with altered checkpoint signaling.

(A). Immunoblot analysis of control or Ras^G12V-infected SXPV/SXPVη cells for indicated checkpoint proteins. All three biological replicates are shown. (B). Representative crystal violet staining of control-infected or Ras^G12V-infected SXPV/SXPVη cells at Day 10. Each cell line was selected with puromycin for 2 days after retroviral infection, followed by seeding at 1000 cells per well (in triplicate). Media was replaced every 2–3 days. After 10 days, colonies were fixed with methanol and stained with crystal violet. (C). Quantitation of survival in Ras^G12V-infected SXPV/SXPVη cells at Day 10, relative to respective controls. The intensity of crystal violet staining in each well was quantified using 590 nm absorbance after destaining. Raw values/well were normalized to a control well with no cells. Survival percentages were calculated as the ratio of normalized mean values from Ras^G12V-infected wells to mean values from control-infected wells. Data represent mean +/- SEM of five biological replicates.