Figure 6
There is more active cPKC in MDA-MB231 cells as compared to MCF-7 cells as determined by reactivity with anti-C2Cat antibodies.
(A) MDA-MB-231 and MCF-7 cells were treated or non-treated with PMA as described above in legend to Fig. 5B. Cell lysates were prepared, fractionated and analyzed by Western blot. The amount of α and γ PKC present in the particulate fraction (active PKC) relative to the total amount of kinase was determined by probing for PKCα and γ with anti-V5 domain antibodies. Quantitative analysis and statistical significance determined by ANOVA-Bonferroni test (n = 3) where **p < 0.01 and *p < 0.05 (B) Immunoprecipitation of active PKCα and γ, by anti-C2Cat, from MDA-MB-231 and MCF-7 cells treated or non-treated with PMA as described above. Lysates were prepared and immunoprecipitated with anti-C2Cat and probed for PKCα and γ using commercially available rabbit polyclonal antibodies against isoenzyme specific V5 domains (representative experiment of n = 3) Quantitative analysis and statistical significance determined by ANOVA-Bonferroni test (n = 3) where **p < 0.01.
