Alexa fluor 594 red goat anti mouse igg

Huh7 cells were seeded onto 24-well plates and transfected with pcDNA3.1-HCV core, pcDNA3.1-GBV-B core, or mock plasmid by Lipofectamine 2000 (Invitrogen, Guangzhou, China). The monoclonal antibody (mAb) to HCV core (C1F5 clone) or GBV-B core (1E5 clone) was used as primary antibody provided in the laboratory (Li et al., 2014 (link)), whereas Alexa Fluor 594 (red) goat anti-mouse IgG (Invitrogen) was used as secondary antibody for detection of the core protein in transfected cells. Diamidinophenylindoldiacetate (DAPI) was added to stain cell nuclei.

Adherent and non-adherent PC3 or PC3/shMcl-1 cells were harvested after a 24 h treatment with 1198, BA, 1198 + BA, or control, applied to slides by smearing, air dried, fixed in formalin for 10 min, permeabilized with 0.1% Triton X-100/PBS for 10 min, rinsed with water, and blocked with goat serum (Vector Laboratories) for 30 min. For DIF, we simultaneously immunostained with rabbit polyclonal Mcl-1 (1/150 dilution) and mouse monoclonal γH2AX (1/200 dilution) for 1 h followed by secondary antibodies Alexa Fluor 488 (green) goat anti-rabbit IgG and Alexa Fluor 594 (red) goat anti-mouse IgG (1/500 dilution; Invitrogen) for 1 h. Mounting medium with DAPI was from Vector Laboratories. Color images were acquired using a Nikon Eclipse 90i fluorescence microscope with FITC/Texas Red filters and merged using Adobe Photoshop 7. A similar DIF experiment was done with PC3/shGFP and PC3/shMcl-1 cells treated with Dox for 4 h.