Figure 2
From: The C-terminal amyloidogenic peptide contributes to self-assembly of Avibirnavirus viral protease
Intracellular assembly of VP4 does not involve other IBDV-encoded proteins or subcellular organelles.
(a) No co-localization of VP4 and IBDV-encoded VP1, VP2 VP3 or VP5 was detected in IBDV-infected cells at 24 hpi. VP1, VP2, VP3 and VP5 in IBDV-infected DF-1 cells were detected with the corresponding mouse mAbs followed by TRITC-conjugated anti-mouse IgG (Red). VP4 was detected by rabbit anti-VP4 antibody followed by FITC-conjugated anti-rabbit IgG (Green). (b) Dot-like VP4 signal partially (white arrow) co-localize with VP2 at 24 hpi in some of IBDV-infected cells. VP2 was detected with mouse anti-VP2 mAb followed by FITC-conjugated anti-mouse IgG (Green). VP4 was detected by rabbit anti-VP4 antibody followed by TRITC-conjugated anti-rabbit IgG (Red). (c) Subcellular relationship of VP4 with subcellular organelles in IBDV-infected cells (MOI = 1) which were pre-transfected with living colors subcellular localization vectors pDsRed2-ER, pDsRed2-Mito, pDsRed2-Peroxi, pEGFP-actin, pEYFP-Golgi or pEYFP-Mem. Cells were fixed and stained with mouse anti-VP4 mAb 24 hpi and FITC/TRITC-conjugated goat anti-mouse IgG. (d) Expression kinetics of VP4 in pCI-wtVP4-transfected DF-1 cells. VP4 was stained at 6, 12 and 24 hpi with anti-VP4 mAb followed by FITC-conjugated anti-mouse IgG. (e) Expression kinetics of EGFP-VP4 protein in pEGFP-wtVP4-transfected DF-1 cells. Rod-shaped and mass-like structures of EGFP-VP4 protein were present in the cytoplasm and nucleus at 6, 12 and 24 h after transfection. Nuclei were stained with DAPI.
