Figure 2

Host protein HSP70-4 was associated with the BmNPV polyhedra matrix. (A) Western blot analysis of HSP70-4 in the BmNPV polyhedra matrix. The fraction over the upper sucrose gradient was loaded for the Western blot analysis. Images were visualized using a chemiluminescence detection system. Lane M, the protein marker. (B) Co-localization of polyhedrin with HSP70-4 in the polyhedra in infected BmN cells. Cells were infected with the BmNPV T3 isolate at an MOI of 10 TCID₅₀/cell. At 48 h p.i. the cells were fixed, permeabilized, and then incubated in a buffer containing mouse monoclonal anti-polyhedrin and rabbit polyclonal anti-HSC/HSP70 antibodies. Next, the cells were stained with the Texas Red-conjugated goat anti-mouse IgG, FITC-conjugated goat anti-rabbit IgG and Hoechst 33342, washed with 1× PBS, and then imaged under a Leica TCS SP5 confocal laser scanning microscope using the Texas Red filter set for the Texas Red-conjugated antibody and the FITC filter set for the FITC-conjugated antibody. In the merged images, yellow suggested an association between polyhedrin and HSP70-4 in the polyhedra matrix (Merge I), and magenta indicated the nuclear location of the polyhedra (Merge II).