FIG. 3.

Activity of specific PKC isoforms in L6 cells transfected with IRS-1 rib. (Top) L6hIR2 cells transfected with either c-Rib or IRS1-Rib were stimulated with 100 nM insulin for 30 min, lysed, and precipitated with antibodies (Ab) to specific PKC isoforms as indicated. PKC activity was then assayed in the immunoprecipitates as described in Materials and Methods. Bars represent the means ± standard deviations of duplicate determinations in three independent experiments. (Bottom) For a control, lysates from the L6-3F2 cells and from the L6hIR2 cells (either those which were not transfected with ribozymes or those transfected with c-Rib or IRS1-Rib) were separated by SDS-PAGE and Western blotted with specific antibodies toward the PKC isoforms as indicated. Blots were revealed by ECL as described in Materials and Methods. The autoradiograph shown is representative of three independent experiments.