Figure 4.

Identification of the interaction between Nsp2 and the host proteins ATP1B3, DNAJA1, and ISCA1. (A). Colocalization analysis between Nsp2 and the host proteins ATP1B3, DNAJA1, and ISCA1. DF-1 cells were cotransfected with pEGFP-Nsp2 and pDsRed-tagged ATP1B3, DNAJA1, or ISCA1 for 36 h, and the nuclei were stained with DAPI before observation using confocal microscopy. (B–G) Co-IP analysis to identify Nsp2 interaction with host proteins. HEK293T cells were co-transfected with pcDNA3.1-Nsp2-Flag and Myc-tagged ATP1B3, DNAJA1, or ISCA1. After 36 h, cellular protein samples were collected and separately incubated with anti-Flag magnetic Beads, anti-Myc magnetic Beads, and IgG magnetic beads. The eluted proteins were then separated by SDS-PAGE and analyzed by Western Blotting using specific primary and secondary antibodies.