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. 2014 Jan 9;7(1):46–57. doi: 10.3390/ph7010046

Figure 4.

Figure 4

Protein expression analysis of RB-E2F-1 interaction and EMSA in PANC-1 cells: (A) 10 µg protein extracts of control and ritonavir-treated cells were resolved by SDS-PAGE, transferred to a nitrocellulose membrane, and probed with RB, CDK4, and p21 as indicated. β actin was used as a loading control. p-RB: hyper-phosphorylated retinoblastoma protein; (B) Upper panel: Indicated concentrations of ritonavir-treated cell lysates were immunoprecipitated with E2F-1 polyclonal antibody and probed with RB monoclonal antibody. Middle panel: Western blot analysis for E2F-1 in cells treated with indicated ritonavir concentrations. Bottom panel: control β actin levels; (C) EMSA with E2F-1 consensus binding site. Lane 1: Control nuclear extract; Lane 2: Nuclear extracts of lentiviral E2F-1 knockdown PANC-1 cell line; 3: Nuclear extracts of 25 µM ritonavir-treated cells.