Immortalized <i>FHL2^−/−</i> MEFs stably expressing cyclin D1 are susceptible to transformation by oncogenic Ras.

<p>(A) Increased cyclin D1 expression in immortalized <i>FHL2^−/−</i> 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 <i>cyclin D1-HA/FHL2^−/−</i> 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 <i>cyclin D1-HA/FHL2^−/−</i> cell lines transduced with H-RasV12. (D) Western blot analysis of p53 induction by doxorubicin in the indicated cell lines as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0003761#pone-0003761-g003" target="_blank">Fig. 3A</a>.