Fig. 1.
Nidovirus replication in cyclophilin-knockout cell pools. A) Huh7 cells were transduced with lentiviruses expressing PPIA-, PPIB-, PPIC-, or PPID-specific gRNAs (Huh7-CypKO,pool cells) to achieve CRISPR/Cas9-mediated knockout of gene expression. The cell pools were analyzed for CypA, CypB, CypC, and CypD protein expression by Western blot analysis. Transferrin Receptor was used as loading control. B-D) Huh7-CypKO,pool cells were infected with MERS-CoV (B), HCoV-229E (C), or EAV (D) at an MOI of 0.01. Virus yields at 48 h p.i. (B, C) or 32 h p.i. (D) were determined by plaque assay. Results presented are the mean ± SD of triplicate harvests from a representative experiment.