FIG 8.

Binding of G3BP1 and FXR1 to the EEEV nsP3 HVD is mediated by their NTF2-like and Agenet-like domains, respectively. (A and B) Schematic representations of VEEV (A) and SINV (B) replicons encoding the Flag-GFP-HVDeeev fusion protein and schematic representation of full-length and deletion mutant G3BP1-GFP (A) and FXR1-GFP (B) fusions stably expressed in G3bp dKO- and Fxr tKO-based cell lines. (C) The indicated cell lines were infected with packaged VEErep/Flag-GFP-HVDeeev or SINrep/Flag-GFP-HVDeeev replicons at an MOI of 20 inf.u/cell. At 2.5 h p.i., cells were harvested, and after lysis, protein complexes were isolated on magnetic beads loaded with Flag-specific MAbs. The presence of G3BP1 and FXR1 variants in isolated complexes and cell lysates was analyzed by Western blotting using GFP-specific MAbs.