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cmi12066-sup-0001-si.eps1251K

Fig. S1. Protein expression level of stable HEK293 cells.

A. Protein extracts of HEK293 cells stably expressing GFP, GFP-CaeA and GFP-CaeB were separated by SDS-PAGE, transferred to a PVDF membrane and probed with antibodies directed against GFP and actin. Shown is one representative immunoblot out of two independent experiments.

B. HEK293 cells and HEK293 cells stably expressing GFP, GFP-CaeA and GFP-CaeB were analysed by flow cytometry. Depicted is one representative experiment out of at least three independent experiments with similar results.

cmi12066-sup-0002-si.eps3971K

Fig. S2. Inducible expression of reCasp 3 is saturating for triggering apoptosis in GFP- and GFP-CaeB-expressing cells. HEK293 cells stably expressing GFP and GFP-CaeB were transfected with pTet-On regulator plasmid and indicated concentrations of pTRE response plasmid encoding activated caspase 3. Five hours post transfection, cells were either left untreated or treated with doxycycline (1 μg ml−1) to induce protein expression. After 18 h at 37°C in 5% CO2 protein extracts were separated by SDS-PAGE, transferred to a PVDF membrane and probed with antibodies against cleaved PARP or actin. Depicted is one representative immunoblot out of two independent experiments with similar results.

cmi12066-sup-0003-si.eps784K

Fig. S3. CaeB reside in the mitochondrial fraction. HEK293 cells stably expressing GFP and GFP-CaeB were lysed, homogenized and mitochondrial and cytosolic fractions were separated. Both fractions were separated by SDS-PAGE, transferred to a PVDF membrane and probed with an anti-GFP antibody. Shown is one representative immunoblot out of at least three independent experiments with similar results.

cmi12066-sup-0004-si.doc39K

Table S1. Plasmids and primers used in this study.

Table S2. Primers used in this study.

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