PKC in erythrocytes. (A) Translocation of PKC activity from cytosol to membrane in uninfected erythrocytes exposed to DMSO (□) or 500 nmol/L PMA (▨) for 15 minutes. PKC activity given is the total kinase activity minus activity measured in the absence of lipid activators (PMA/phosphatidyl-serine/Triton-X–100 mixed micelles) and the presence of 20 μmol/L PKC.19-36 Activities represent the mean of duplicate assays + range. This result is representative of six separate experiments. (B) Western blot of whole cell lysates from uninfected cells, prepared and separated on a 10% acrylamide gel (107 cell equivalents/lane) as described in Materials and Methods. Blots were incubated with monoclonal antibodies as follows: lane 1, anti-PKCα at 0.05 μg/mL; lane 2, anti-PKCβ at 0.16 μg/mL; lane 3, anti-PKCγ at 0.25 μg/mL; lane 4, anti-PKCε at 0.5 μg/mL; lane 5, anti-PKCθ at 1 μg/mL; lane 6, anti-PKC ι at 1.6 μg/mL and lane 7, anti-PKCλ at 1.6 μg/mL. No bands were detected in parallel blots incubated with control antibodies or second layer only (not shown).
