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. 2004 Dec;78(23):12747–12761. doi: 10.1128/JVI.78.23.12747-12761.2004

FIG. 7.

FIG. 7.

PKR activation is impaired in dsRNA treated PERK−/− MEFs. (A) PERK+/+ and PERK−/− MEFs were transfected with dsRNA (10 μg/ml, lanes 2 and 4) or mock transfected (lanes 1 and 3) or treated with TG (1 μM, 2 h; lanes 5 and 6). Proteins extracts were harvested at the indicated times and subjected to immunoblot analysis with the serine 51 phospho-specific eIF2α antibody (top panel) or with the eIF2α panspecific antibody (lower panel). (B) Protein extracts (200 μg) from dsRNA transfected PERK+/+ and PERK−/− MEFs were immunoprecipitated with a rabbit polyclonal anti-mPKR antibody (D-20) and subjected to phosphorylation in vitro in the presence of [γ-32P]ATP. The immunoprecipitated PKR was subjected to SDS-10% PAGE and autoradiography (top panel). Protein extracts (50 μg) were subjected to immunoblot analysis with a rabbit polyclonal phosphothreonine 446 PKR antibody (second panel) or a mouse monoclonal actin antibody (bottom panel).