Figure 5.

siRNA1 and siRNA2 induce a decrease in AKAP350 expression. HeLa cells were transfected with RNA duplexes targeted to two different AKAP350 specific sequences (siRNA1 and siRNA2) or with the RNA duplexes synthesized using a scrambled sequence of the siRNA1 target sequence (siRNA control) and allowed to grow for 48 h. The cells were scraped and lysed in the presence of protease inhibitors. The samples were resolved by SDS-PAGE and transferred to nitrocellulose or Immobilon membranes for the analysis of AKAP350 or CIP4 expression. The densitometry of the bands indicated a reduction of total AKAP350 protein to 20% of control with the siRNA1 and to 40% of control with the siRNA2 transfected cells. The RNA interference for AKAP350 expression did not modify the CIP4 protein levels.