Expression pattern of PKZ 12 h after induction by poly(I:C). RT-PCR was performed on the indicated tissues, by using primers covering the complete ORF. The constitutively expressed receptor for activated C kinase 1 (RACK1) is shown as a loading control at the bottom. Expression patterns in tissues from untreated animals are in Fig. 9.

Inhibition of protein synthesis by PKZ. (A) HEK293T and CHO cells were cotransfected with 2.5 μg of the luciferase plasmid pGL3promoter and 0.5 μg of the expression vector pcDNA3.1 containing the indicated inserts. Luciferase activity was normalized for protein content. A control experiment by using the empty pcDNA3.1 vector was used as 100%. An asterisk denotes the position in the constructs of the K199R mutation that abolishes kinase activity. Each bar is the average of three independent experiments. (B) Western blot analysis of HEK293T cells transfected with the expression constructs shown in A. His-tagged proteins were detected by an anti-His-tag antibody. The asterisk marks a nonspecific band detected in all lanes. Silver staining demonstrated comparable loading of proteins (data not shown).

Expression of drPKZ in tissues from untreated animals. Splice variants A–D are marked on the right. Lower shows the expression of the constitutively expressed receptor for activated C kinase 1 (RACK1) as a control.