FIG 9.

Colocalizations of TSWV G_N with endoCP-G_N or cyclophilin in insect cells. Open reading frames of cyclophilin, endoCP-G_N, and TSWV G_N were cloned into Drosophila Gateway vectors (under the control of the Hsp70 promoter), and the resulting pHWR and pHWG expression plasmids were used for the following fusion proteins: cyclophilin-RFP, endoCP-G_N-RFP, and TSWV G_N-GFP. The recombinant plasmids, pHWR-cyclophilin, pHWR-endoCP-G_N, and pHWG-TSWV-G_N, were singly transfected or cotransfected into insect Sf9 cells. All transfection reactions were performed using the Cellfectin II reagent. Mock, no DNA treatment (top left panels) and cotransfection of pHRW and pHGW expression plasmids (bottom left panels) were used as controls. Cells were stained with DAPI 72 h posttransfection and then visualized using the Cytation 5 cell imaging multimode reader (BioTek, Winooski, VT) to detect red and green fluorescence. Exposure settings (LED intensity/integration time/camera gain) of the mock control were set as the baseline parameters for the analysis of all other treatments. Cells were visualized with a 40× objective. Scale bars = 10 μm.