Gtx109753

Vero E6 or Sf21 cells were seeded onto an 8-well Millicell^® EZ slide (4 ×10⁴/well), and the cells were transduced with recombinant baculovirus. The cells were fixed and stained as described previously [4 (link)]. Briefly, the cells were fixed with 4% paraformaldehyde, and then permeabilized with 100% acetone at -20°C. After blocking with 3% BSA for 1 h, the cells were then incubated with primary antibody overnight at 4°C. Primary antibodies anti-IE2 antiserum (1:10000), anti-His antibodies (1:10000, Abcam ab18184), anti-FLAG (1:500, Sigma F1804), anti-HSP70 (1:5000, Genetex GTX111088) or anti-HSP90 alpha (1:2000, Genetex GTX109753) were diluted in blocking buffer. After incubation overnight, the cells were washed three times with DPBST (DPBS containing 0.1% Tween 20) and then incubated for 1 h with secondary antibodies including Dylight goat 549 anti-rabbit IgG, Dylight 405 goat ani-mouse IgG (1:200, Jackson) or Alexa Fluor 488 goat anti-mouse IgG (1:500; Life Technologies). Also, Alexa Fluor 488-conjugated DNaseI (1:500; Life Technologies) was used for G-actin detection, and Hoechst dye (Life Technologies) for nucleus staining. After washing three times with DPBST, cells were sealed with mounting solution (Fluoromount-G, SouthernBiotech). Images were collected using a Zeiss laser confocal microscope (LSM 780).