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. 2010 Dec 14;30(2):379–394. doi: 10.1038/emboj.2010.326

Figure 7.

Figure 7

Coimmunoprecipitation of A-SMase with caspase-8, caspase-7, cleaved caspase-7 and caspase-3. (A) HeLa cells stably transfected with GFP-tagged A-SMase were lysed and anti-GFP (Invitrogen, Molecular Probes), anti-caspase-8 (Santa Cruz), anti-caspase-7 (Abcam) and anti-caspase-3 (Cell signaling) antibodies were used for precipitation. Western blots performed with anti-GFP (Invitrogen, Molecular Probes), anti-caspase-8 (Santa Cruz), anti-caspase-7 (Abcam), anti-cleaved-caspase-7 (Cell signaling) and anti-caspase-3 (Cell signaling) antibodies demonstrate that caspase-7 coprecipitates with pro-A-SMase-GFP and vice versa in contrast to caspase-8 and caspase-3, which are neither precipitated with anti-GFP nor with anti-caspase-7 antibodies (in the case of caspase-8). (B) Immunoprecipitation of endogenous A-SMase with the anti-A-SMase antibody and of caspase-7 with anti-caspase-7 antibodies (Abcam) from HeLa wild-type cell lysates. Western blots show that caspase-7 coimmunoprecipitates with A-SMase and vice versa, in line with the results shown in (A). Figure assembled from cropped lanes of the same blots of the respective antibody (see primary scans in Supplementary data).