The effect of PKC- and PKC-β2 ribozymes on CD34⁺ cells cultured with EPO and SCF.

The CD34⁺ cells were incubated at 30 000 cells/well in flat-bottomed, 96 well plates with PKC-α ribozyme (2 μmol/L), PKC-β2 ribozyme (2 μmol/L), or medium for 24 hours and then stimulated with EPO (5 U/mL) and SCF (50 ng/mL.). (A) Cells were stained with the indicated mAbs 8 days after transfection with ribozymes. (B) Cytosolic proteins were prepared from cells cultured for 6 days after transfection. (C) Cytosolic proteins were prepared from cells cultured for 6 days after transfection. Control cells were transfected with a PKC-ε ribozyme that was not catalytic active. (D) RT-PCR was performed on RNA prepared from cells cultured for 6 days after transfection. PKC-α, 320 bp; actin, 620 bp.