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. Author manuscript; available in PMC: 2014 Jun 1.
Published in final edited form as: Mol Cancer Res. 2013 Feb 28;11(6):638–650. doi: 10.1158/1541-7786.MCR-12-0634-T

Figure 1. High exogenous levels of KRASV12, comparable to endogenous levels found in mutant KRAS NSCLC cell lines, increases transformation of HBECs and induces senescence, which is largely bypassed with p53 knockdown.

Figure 1

A) Immunoblot for KRAS protein expression in HBEC3 cells infected with KRASV12 using either a moderately-expressing retroviral (pBabe-hyg-KRASV12) or high-expressing lentiviral (pL6-KRASV12) vector. Actin was used as loading control. B) Anchorage-independent (soft agar) colony formation in HBEC3 with high (lentiviral) or moderate (retroviral) levels of KRASV12 in the background of both wildtype p53 and p53 knockdown (sh-p53) (t-test). C) Immunoblot of HBEC3p53,KRAS soft agar clones confirming p53 and KRASV12 manipulations. The presence (+) or absence (−) of each manipulation is shown. D) Anchorage-dependent (liquid) colony formation ability of HBEC3p53,KRAS soft agar clones. E) Quantification of SA-β-gal staining found KRASV12-induced senescence in HBEC3 cells was significantly lower in cells with p53 knockdown compared with p53 wildtype (t-test). F) Anchorage-dependent colony formation assay to compare acute KRASV12-induced toxicity in HBEC3 and HBEC4 with wildtype p53 or p53 knockdown. G) Immunoblot of HBEC3 cell lysates harvested seven days after infection with KRASV12 or LacZ lentivirus. *P < 0.05, **P < 0.01, ***P < 0.001. Full-length blots are presented in Supplementary Figure S8).