Figure 6. Immortalized FHL2^−/− MEFs stably expressing cyclin D1 are susceptible to transformation by oncogenic Ras.

(A) Increased cyclin D1 expression in immortalized FHL2^−/− MEFs after introduction of cyclin D1-HA transgene was confirmed by immunoblotting with anti-cyclin D1 and anti-HA antibodies. (B) Representative phase contrast images of immortalized wt and cyclin D1-HA/FHL2^−/− cell lines transduced with H-RasV12 or empty vector. (C) Soft agar assay. Cells were initially seeded at 5×10⁴ cells/plate and cultured for 10 days. Images are representative of three wt and two cyclin D1-HA/FHL2^−/− cell lines transduced with H-RasV12. (D) Western blot analysis of p53 induction by doxorubicin in the indicated cell lines as described in Fig. 3A.