Figure 6.

Reduction of AKAP350A expression by RNA interference alters CIP4 distribution and induces vesiculation of the Golgi apparatus. HeLa cells were transfected with the AKAP350A-targeted RNA duplexes siRNA1 and siRNA2 or with the siRNA control, and allowed to grow for 72 h. After fixation, cells were dual stained with the 14G2 anti-AKAP350 and r-CIP4 antibodies (first column), anti-AKAP350A and anti-p58 (second column), anti-AKAP350A and anti-golgin-97 (third column), and anti-AKAP350A and anti-α-tubulin (fourth column). Approximately 90% of the cells transfected with siRNA1 and siRNA2 showed a decrease in AKAP350A staining that was more prominent in the cells expressing the siRNA1. Approximately 40% of the siRNA1 and 20% of the siRNA2 cells showed no AKAP350A staining. The reduction in AKAP350 expression was confirmed by Western blot (see Figure 5). The viability of the cells was confirmed by evaluation of the DAPI staining and the phase contrast images of the cells (our unpublished data). Bar, 2.5 μm.