Figure 1.

Protein Synthesis and Accumulation in Murine Cancer Cells Infected with D9- or D10-Deficient VACV

(A) Murine MBT2 bladder carcinoma, murine 4T1 breast carcinoma, or NHDFs were mock-infected (mock) or infected (MOI = 3) with WT VACV, D9-deficient VACV (ΔD9), or D10-deficient VACV (ΔD10). At 18 hours post-infection (hpi), cells were metabolically pulse labeled with [³⁵S]Met-Cys for 30 min. Total protein was collected and separated by SDS-PAGE, and [³⁵S]-labeled proteins were visualized by exposing the fixed, dried gel to X-ray film. Molecular mass standards (in kDa) are shown on the left. Representative radiolabeled proteins in mock-infected NHDFs that decrease in infected cells (consistent with host shut-off) are indicated (⋅). Representative radiolabeled proteins in mock-infected MBT2 or 4T1 cells that persist in infected cells are indicated (o). (B) Samples in (A) were analyzed by immunoblotting using anti-VACV polyclonal antisera as described.⁵⁹