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. 2002 Mar 15;109(6):827–836. doi: 10.1172/JCI14146

Figure 5.

Figure 5

Caspase-9 mediates the late Cer accumulation from the SM cycle. (a and b) LNCaP cells were cultured treated with the complex indicated (10 μM etoposide, 10 μM Rottlerin, 50 μM FB-1) for 9 hours, and cell lysates were analyzed by Western blot analysis using anti–caspase-9 Ab and anti–caspase-3 Ab (a), and caspase-9 protease activity was assayed as described in Methods (b). Bars represent SD. Experiments were repeated twice in triplicate with similar results. (c) LNCaP cells were treated with the complex indicated (10 μM etoposide, LEHD-fmk, DEVD-CHO, zVAD-fmk) for 9 hours, and Cer levels (upper panel) and SMase activity (graph) were measured. Cer data are expressed as fold increase relative to the control and are representative of three experiments. Bars represent SD. Experiments were repeated three times with similar results. (d) LNCaP cells were treated with the complex indicated (10 μM etoposide, 50 μM LEHD-fmk, 100 μM DEVD-CHO) for 1 hour or 9 hours, and mitochondrial fractions were analyzed using anti-PKCδ Ab.