FIG 3.

Molecular characterization of SK6 DNAJC14-KO cells. (A) Naïve SK6 cells and SK6 DNAJC14-KO cell clones were infected with CSFV (Alfort-Tübingen) at an MOI of 3 and analyzed by NS3-specific IF assay at 72 hpi (top). (Lower) Cell nuclei were visualized by DAPI staining. (B, top) Scheme of genomic PCR amplification with the intron-specific primer flanking exon 4 of the porcine DNAJC14 genomic DNA. Analysis of PCR amplicons obtained from PCR amplification of genomic DNA isolated from naive SK6 cells (2 amplicons, SK6 naive 1 and 2) and SK6 DNAJC14-KO cell clone 9 (12 amplicons, SK6 DNAJC14-KO amplicons 1 to 12) is shown. Sequences surrounding the guide RNA-mediated genome alteration of the DNAJC14 gene are shown. (Middle) Porcine DNAJC14 genomic DNA sequence is shown as a reference. Porcine DNAJC14 Jiv90 DNA and amino acid sequence is depicted for naive SK6 cells and SK6 DNAJC14-KO cell clone 9 with the two identified guide RNA-mediated DNAJC14 genome alterations (Δ22 and Δ6). (Lower) Depicted amino acid sequence starts with the beginning of the Jiv90 domain. Positions of the stop codons resulting from deletions introduced by the DNAJC14 guide RNA are indicated by a star.