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. 2022 Apr 27;13:868377. doi: 10.3389/fmicb.2022.868377

FIGURE 2.

FIGURE 2

Substitution of residues s1–s6 of ALV-A with ALV-E reduced the Tva binding affinity and infectivity of DF-1 cells. (A) Schematic representations of the replacement fragments s1–s6 in hr1. (B) DF-1 cells were incubated with chimeric gp85 protein s1–s6 and subsequently infected with RCASBP (A)-EGFP supernatants. The percentage of GFP-positive cells was measured by FACS for blocking analysis of the binding of chimeric gp85 protein to Tva. (C) The interaction of chimeric gp85 proteins s1–s6 with Tva-HA-Fc. (D) The gray scale value of the protein band was quantified using Image Studio Lite Version 5.2, and gp85 (IP)/Tva (IP) was calculated. (E) DF-1 cells transfected with recombinant RCASBP (A/E)-s1-s6-EGFP vectors were visualized under a fluorescence microscope, and the percentage of GFP-positive cells (indicated with red color letters) was detected by FACS 7 days posttransfection. *P < 0.05, **P < 0.01, ****P < 0.0001.