Intra-Lesional Injection of the Novel PKC Activator EBC-46 Rapidly Ablates Tumors in Mouse Models
A. The structure of EBC-46. Structure of PMA shown as a comparison. B. Production of reactive oxygen species following treatment of PMN cells. PMN cells were pre-loaded with dihydroethidium bromide and pre-treated where indicated with 1 µM bisindolylmaleimide-l for 15 min, then stimulated with 175 nM (100 ng/ml) of either EBC-46 (left) or PMA (right) for 15 min at 37°C. Untreated PMN cells – red, PMN cells pre-loaded – blue, loaded PMN cells treated with either 175 nM (100 ng/ml) PMA or EBC-46 – green, loaded PMN cells treated with either 175 nM (100 ng/ml) PMA or EBC-46 in presence of bisindolylmaleimide-l – yellow. Data is representative of three independent experiments. C. PKC-EGFP isoform translocation in transiently transfected HeLa cells following 1 h treatment with either 175 nM (100 ng/ml) PMA or EBC-46. Data is from assessment of at least 50 cells per well from each of triplicate transient transfection experiments. Error bars – SD. D. HeLa cells were treated with either 175 nM (100 ng/ml) EBC-46 or PMA for 1 h, or vehicle alone (Control). PKC-α was detected by immunofluorescence. E. PKC kinase assay. HeLa cells were treated with the indicated amounts of EBC-46 or PMA for 1 h. A total of 30 µg of lysate was used to determine PKC-specific kinase activity. Assays were performed in triplicate (n = 3) with the mean ± SD shown.