FIG. 5.

Coexpression of wt PKCα augments V14Rho-enhanced AP-1 transcriptional activity, while coexpression of N-PKCα blocks activated-Rho-plus-PMA-enhanced AP-1 transcriptional activity. Jurkat cells were cotransfected with 10 μg of reporter construct and 10 μg of V14Rho and/or pEBB-wt PKCα (A) or 10 μg of V14Rho plus 10 μg of pEBB or a construct encoding N-PKCα (B). Cells were left unstimulated or stimulated with PMA (10 ng/ml) for 7 h. AP-1 luciferase activity was determined. (C and D) V14Rho (lane 1)- and N-PKCα construct (lane 2)-transfected cells (5 × 10⁶) were lysed, immunoprecipitated with glutathione-Sepharose beads, and immunoblotted with anti-GST (C) and anti-c-Myc (9E10) (D) antibodies.