Inhibition of Translation Initiation by Protein 169: A Vaccinia Virus Strategy to Suppress Innate and Adaptive Immunity and Alter Virus Virulence
Fig 6
Puromycin labeling of nascent polypeptides.
(A, B) HEK 293 Trex 169 (A) or C6.TAP (B) cells were uninduced or induced with DOXY (1 μg/ml) for the indicated times. Nascent proteins were then labeled with 5 μg/ml of puromycin for 25 min. Cell lysates were normalized by BCA assay and were resolved by SDS-PAGE and analyzed by immunoblotting using the indicated antibodies. Data shown are one representative experiment (n = 2). (C) HeLa cells were either mock-infected or infected with v169 and vΔ169 at 5 PFU/cell for the indicated times. The samples were processed as described for (A, B) above. Data shown are from one representative experiment (n = 2). The positions of molecular size markers in kDa are indicated on the left for all panels.