Western blot using affinity purified anti-ROCK-2 pY256 antibody shows detection of phosphorylated ROCK-2 in transfected 293T cells. Lane 1 shows lysate expressing exogenous ROCK-2 where the insert contains the transversion Y256A. Lane 2 shows the lysate expressing WT ROCK-2. A lysate containing a GST null vector also shows no specific staining (data not shown). Primary antibody was used at a 1:1000 dilution. Personal Communication, A. Papageorge and X. Qian, NCI, Bethesda, MD.