FIG. 2.
Protein synthesis in HCMV-infected cells following thapsigargin and CLT treatment. Thapsigargin (black bars), CLT (gray bars), and DMSO (open bars) were added to HCMV-infected HFF cultures during the indicated 24-h periods after infection (6 to 30, 24 to 48, and 48 to 72 hpi). Mock-infected cells were similarly treated for 24 h. At the end of the 24-h treatment periods, the cells were pulse labeled with [35S]methionine-[35S]cysteine for 30 min, and total protein extracts were prepared to measure protein synthetic levels. (A) Total protein was TCA precipitated from each sample; the bars indicate the levels of 35S incorporation relative to that of the DMSO-treated control (set at 100%). (B) The level of [35S]methionine-[35S]cysteine incorporation into HCMV IE1/IEP72 in each sample was evaluated by immunoprecipitation from lysates, which was followed by SDS-PAGE and quantitation of the IE1/IEP72 bands by PhosphorImager analysis.