Transcript and protein levels of PRKCZ in parental and <i>PRKCZ</i>-transfected SKOV3 and OVCAR cell lines.

<p>(A, B) The endogenous transcript and protein levels of PRKCZ are low in both SKOV3 and OVCAR3 cells, as determined by real-time PCR and western blot analysis. Human acute monocytic leukemia cell line THP-1, which expresses a high level of PRKCZ, was used as a positive control. Beta-actin protein was probed on the blot as a loading control. The beta-actin loading control is also shown (lower panel). (C-E) Comparison of PRKCZ transcript and protein expression in SKOV3 parental control (PC), control vector (N2), and a selection of PRKCZ clones by quantitative real-time RT-PCR, western blot analysis with PRKCZ-antibody, and fluorescence microscopy. (F-H) Comparison of PRKCZ transcript and protein expression in OVCAR3 parental cells (PC), control vector (N2), and a selection of PRKCZ clones by quantitative real-time RT-PCR, western blot analyses with PRKCZ-antibody, and fluorescence microscopy. For both SKOV3 and OVCAR3, a band of ~95 kDa was detected by western blotting in stable cell lines that over-express PRKCZ, which corresponded with molecular weight of GFP-tagged PRKCZ protein. A faint band of ~67 kDa corresponded to endogenous PRKCZ. The three PRKCZ clones for both SKOV3 and OVCAR3 were selected for further experiments.