Fig. 3.

Interaction with the KDELR is important for the viral role of coatomer. (A) HeLa cells, either treated with a combination of siRNAs that targets all human KDELR genes for 48 h or mock-treated, were infected with WR virus for 24 h. The vaccinia replication assay was then performed, with the mean and standard error from 3 experiments shown (P < 0.01). Gel shows protein levels in whole-cell lysate by immunoblotting for proteins indicated. (B) HeLa cells, either treated with siRNAs against individual KDELR genes as indicated for 48 h or mock-treated, were infected with WR virus for 24 h. The vaccinia replication assay was then performed, with the mean and standard error from 3 experiments shown. (C) HeLa cells, stably transfected with either a siRNA-resistant form of wild-type KDELR1 or a mutant KDELR1 defective in binding to coatomer or untransfected as control, were treated with siRNA against native KDELR1 for 48 h. Cells were then infected with WR virus for 24 h. The vaccinia replication assay was then performed to obtain a mean with standard error from 3 experiments. Values are normalized to that of control cells. (D) HeLa cells treated with siRNA against KDELR1, stably expressing wild-type KDELR1, mutant KDELR1, or untransfected as control, were infected with virus for 24 h. Coatomer was then immunoprecipitated from cells from the different conditions followed by immunoblotting for the proteins indicated.