Fig. 1. Silencing of SV40 large T antigen in a stably transformed rat fibroblast cell line (F5). Triple fluorescence staining of cells transfected with T-antigen siRNA duplex (A,C,E), and with GL2 luciferase siRNA duplex serving as control (B,D,F). (A,B) Staining with T-antigen specific antibody, (C,D) staining with NuMA specific antibody, (E,F) Hoechst staining of chromatin. The few cells cells in A that are stained for T antigen were probably nontransfected. (G) Western blot of cells transfected with T antigen siRNA (left) or luciferase siRNA (right) probed with T antigen-specific antibody. The blot was stripped and reprobed with vimentin antibody (bottom). Magnification 480x (A-F).