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. 2000 Jul 5;97(14):7748–7753. doi: 10.1073/pnas.97.14.7748

Figure 2.

Figure 2

A dominant genetic determinant is important for RB signaling. (A) The C-C, S-S, and C-S cell fusions were transfected with the E2F promoter reporter plasmid E2F/Luc, CMV-β-galactosidase (β-gal), and vector or PSM-RB. (B) The C-C, S-S, and C-S cell fusions were transfected with the cyclin A promoter reporter plasmid −608Luc, CMV-β-gal, and either vector or PSM-RB. (C) Fusion cells were cotransfected with H2B-GFP and either CMVNeoBam (lanes 1, 3, and 5) or PSM-RB (lanes 2, 4, and 6). Forty-eight hours after transfection, the transfected cells were sorted from the untransfected cells by fluorescence-activated cell sorting for GFP fluorescence. Lysates were prepared from these cells, and protein was resolved by SDS/PAGE and then immunoblotted for cyclin A, Cdk2, cyclin E, and Cdk4 proteins. (D) The C-C, S-S, and C-S cell fusions were cotransfected with a H2B-GFP expression plasmid and either vector or PSM-RB. BrdUrd was added 48 h after transfection and cells stained for BrdUrd incorporation. The displayed values were determined from two independent experiments with at least 150 transfected (GFP positive; solid bars) or 150 untransfected (GFP negative; open bars) cells counted per experiment. (E) The C-C cell fusions were cotransfected with H2B-GFP expression plasmid and either BRG-1 or PSM-RB + BRG-1.