U118 cells treated with control or ARAP2 siRNA were transfected with plasmids for expressing either Flag-ARAP2 or Flag-[R728]ARAP2. The plasmid without cDNA for ARAP2 (vector) were used as a control. Cells were plated on fibronectin coated glass slides for 6 hrs, fixed and stained for pAkt and the Flag epitope, followed by immunofluorescence microscopy. (A) Representative images with the top panel showing staining for the Flag epitope to identify transfected cells and the lower panel staining for pS473Akt. (B) The summary data is shown as the mean±S.E. from quantifying 20-22 cells under each condition from 2 experiments. Scale bar, 20 μm. **, p<0.01, ***, p<0.001.