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. 2011 Feb 22;19(6):1090–1096. doi: 10.1038/mt.2011.17

Figure 3.

Figure 3

GFP-DEVD-ssGluc is cleaved specifically in response to apoptosis. (a,b) Gli36 cells-expressing GFP-DEVD-ssGluc were treated with different concentration of doxorubicin in triplicates. Sixteen hours later, cell lysates as well as conditioned medium were analyzed by western blotting using an antibody against (a) Gluc and 50 µl aliquots of conditioned medium were assayed for (b) Gluc activity. (c,d) Gli36 cells-expressing GFP-DEVD-ssGluc were treated with either different amounts of doxorubicin in triplicates, or a combination of both doxorubicin and ZVAD-fmk, a caspase inhibitor. Sixteen hours later, cell lysates and conditioned medium were analyzed by western blotting for Gluc as well as caspase-3 using (c) specific antibodies and conditioned medium was assayed for (d) Gluc activity. Western blots shown are representative from a single well of different groups. For Gluc assay, data presented as mean ± SD for three biological and three experimental replicates.