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. Author manuscript; available in PMC: 2007 Apr 24.
Published in final edited form as: J Biol Chem. 2005 Apr 18;280(24):23262–23272. doi: 10.1074/jbc.M503701200

FIG.1.

FIG.1

TPA-mediated apoptosis in prostate cancer cell line CWR22-Rv1. A, time course for TPA-mediated apoptosis induction. Apoptosis was measured by bis-benzimide staining at the indicated times. B, FB1 inhibits TPA-mediated apoptosis. Cells were pretreated with 40 μM FB1 for 3 h before 10 ng/ml TPA was added. Apoptotic nuclei were counted after 48- and 72-h incubation by bis-benzimide staining. C, ceramide synthase in TPA-treated CWR22-Rv1 cells. CS activity was measured after incubating the cells with 10 ng/ml TPA for the indicated time. D, ceramide levels following incubation with 10 ng/ml TPA. Samples of TPA-treated cells were taken at the time points shown. Ceramide levels were measured by the diacylglycerol kinase assay, and the bars represent S.D. values. E, effect of radiation on TPA-mediated apoptosis. Cells were incubated with 300 pg/ml TPA for 16 h before irradiation at 20 Gy. Apoptotic nuclei were counted using bis-benzimide staining 72 h after irradiation. This dose of TPA that alone was insufficient to induce significant apoptosis conferred radiation sensitivity (p < 0.01; stratified rank sum) in CWR22-Rv1 cells. The bars represent S.D. values. All studies were repeated at least twice.