Figure 3

Co-localization of HTNV N protein with intracellular scFvs. A) Co-localization of recombinant N protein with intracellular scFvs, COS-7 cells were co-transfected with pEF-N76118 and a intracellular scFv expressing plasmid [(I) L13F3 scFv-CYTO, (II) H34-scFv-CYTO, (III) L13F3-scFv-ER, (IV) H34-scFv-ER], 2 days post transfection, slides were acetone fixed, and costained with human anti-N IgG H34 [green, panels 1] or anti-penda-His mAb [red, panels 2]), merged images of the panels 1 and 2 are presented in panels 3; B) Co-localization of Gc with intracellular scFvs, Stably transfected vero E6 cells [(I) L13F3 scFv-CYTO, (II) H34-scFv-CYTO, (III) L13F3-scFv-ER, (IV) H34-scFv-ER] were infected with 76-118 at an MOI of 0.1, and after 6 days slides were acetone fixed, and costained with anti-Gc mAb AH100 [green, panels 1] or anti-N mAb [red, panels 2]), merged images of the panels 1 and 2 are presented in panels 3. C) Co-localization of viral N protein with intracellular scFvs. Intracellular scFv stably transfected vero E6 cells were infected with 76-118 at an MOI of 0.01, [(I) L13F3 scFv-CYTO, (II) H34-scFv-CYTO, (III) L13F3-scFv-ER, (IV) H34-scFv-ER], or with L99 at an MOI of 0.01 [(V) L13F3 scFv-CYTO, (VI) L13F3-scFv-ER], and after 5-6 days slides were acetone fixed, and costained with human anti-N mAb AH34 [green, panels 1] or mouse anti-His tag mAb [red, panels 2]), merged images of the panels 1 and 2 are presented in panels 3. Excitation wavelengths were 488 and 568 nm; Fluorescence images were obtained with confocal microscope using 100 × objectives