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. 2003 Jan;162(1):93–103. doi: 10.1016/S0002-9440(10)63801-1

Figure 5.

Figure 5.

The peptide-based caspase inhibitors Z-VAD-fmk (A) and Z-ASTD-fmk (B) do not inhibit formation of mature EMAP II in hypoxic B16 melanoma cells. Cells were incubated at 1% oxygen in a hypoxic chamber for the times indicated. Increasing concentrations of the inhibitors were applied. Control cells were grown in a normoxic environment. Supernatants of the cultured cells were subjected to Western blot analysis using the EMAP II-specific antiserum. Only the band corresponding to the mature 23-kd EMAP II protein is shown. C: Z-VAD-fmk inhibits cleavage of proEMAP II/p43 in apoptotic B16 melanoma cells. Apoptosis was induced in B16 melanoma cells and where indicated, Z-VAD-fmk was added at increasing concentrations. The culture medium was analyzed for EMAP II protein by Western blotting. D: Western blot analysis of total lysates of B16 melanoma cells. After stimulation as indicated, cells were lysed and 20 μg total protein were analyzed by Western blotting using the EMAP II-specific antiserum (upper panel). The membrane was reprobed with a GAPDH-specific antibody to ensure equal protein loading (lower panel).