ICAM-1 activates PKCα but not PKCβII in HMVECLs. A

<p>) At 70% confluence, HMVECLs cells pretreated with 10 ng/ml TNFα to induce ICAM-1 expression. At 24 hrs, the cells were suspended and immunolabeled with anti-ICAM-1 antibodies and examined by flow cytometry for ICAM-1 expression. <b>B–D</b>) HMVECLs were pretreated with TNFα as in panel A. At 24 hrs, the endothelial cells were nonstimulated (NS) or stimulated with a confluent monolayer of anti-ICAM-1-coated beads or the control anti-PECAM-1-coated beads for 20 minutes in <b>B</b> and <b>D</b> or for the times indicated in <b>C</b>. The cells were lysed and the activation of PKCβII and PKCα was examined by western blot with <b>B</b>) anti-phospho-PKCβII^Thr641 or anti-PKCβII or <b>C–D</b>) anti-phospho- PKCα^Thr638 or anti-PKCα. Shown are representative blots. Shown are the mean ± SEM of 3 experiments. NT, nontreated. *, p<0.05 as compared to the nontreated (NT) groups.