FIG. 2.

Both pm142 and pm143 bind to PKR in transfected cells. HeLa cells were transfected with the indicated plasmids, all of which express proteins containing His tags alone or with a biotinylation signal. All transfections, except that in lane 4, also included the plasmid encoding biotin ligase (pCS2+BirA). At 48 h posttransfection cell lysates were prepared and incubated with avidin-agarose and analyzed by immunoblot assay as described in Materials and Methods. (A) Membranes with unbound lysates (top panel) or 20% of the bound samples (bottom panel) were probed with the anti-His antibody. (B) Membranes with unbound lysate (top panel) and 80% of the bound samples (lower panel) were subjected to immunoblot analysis with the PKR antibody.