Fig. 2.
RpL40-dependent translation is transcript-specific. (A) VSV protein synthesis. Total [35S]-methionine-cysteine–labeled cytoplasmic proteins were analyzed by SDS/PAGE and detected by phosphorimager. Transfection of cells with nontargeting (NT) or rpL40-targeting siRNA is indicated. (B) Schematic diagram of bicistronic CrPV IRES construct. Translation of firefly luciferase is cap-dependent, whereas translation of renilla luciferase is driven by the CrPV IRES. (C) Firefly luciferase protein synthesis driven from pFR-CrPV. Firefly luciferase was measured from pFR-CrPV transfected lysates at 12 h after transfection and normalized to luciferase units from NT-treated cells. The results are given as the mean ± SD of three independent experiments each performed in triplicate. (D) Renilla luciferase protein synthesis driven from pFR-CrPV. Luciferase levels were measured and normalized as in C. (E) Poliovirus protein synthesis. Cells were infected and total cytoplasmic proteins were analyzed as in A. (F) Microscopy of cells infected with rabies virus-mCherry. (G) Microscopy of cells infected with measles virus-GFP. (H) Microscopy of cells infected with Newcastle disease virus-GFP.