Restoration of cyclin D1 and cell proliferation in FHL2-/- MEFs by re-expression of FHL2. A, up-regulation of cyclins D1, D2, D3, and E and Rb phosphorylation in FHL2-restored cells. Upper panel, mRNA levels of cyclin D1 and FHL2 in immortalized MEFs of different genotypes. Real time RT-PCR was carried out using either cyclin D1- or FHL2-specific primers on total RNA extracted from indicated cell lines. Their expression was normalized to 18 S RNA. The average and S.D. values for three independent experiments are shown. Data are representative of those from three independent clones for WT and FHL2-/- genotypes and three independent pools of FHL2-restored and pBabe-infected cells. The ratio of cyclin D1/18 S or FHL2/18 S in WT MEFs was arbitrarily set at 1. In the lower panel, cell lysates were analyzed by immunoblotting. The asterisk shows hyperphosphorylated Rb. B, growth curves of immortalized WT, FHL2-/-, pBabe-infected and pBabe-FHL2-infected FHL2-/- MEFs. The data presented are the mean ± S.D. obtained from three independent experiments. Data are representative of growth curves from two independent FHL2-infected MEF pools.