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. 2002 Dec 9;159(5):739–745. doi: 10.1083/jcb.200209004

Figure 2.

Figure 2.

Salt and prodomain dependence of caspase-2 complex. (A) HeLa cell lysates in 20 mM Hepes, pH 7.5, were supplemented with either 150 mM KCl or NaCl as indicated before incubation at 37°C for 60 min. Only selected fractions 2–8 and 19–25 were analyzed by immunoblotting. (B) Lysates from control HEK 293T cells or those transfected with a GFP-tagged prodomain-less caspase-2 expression construct were incubated at 37°C for 1 h and subjected to size exclusion chromatography. Fractions were immunoblotted with the caspase-2 antibody (top) or a GFP antibody (bottom). Note that prodomain-less caspase-2–GFP fusion protein (∼60 kD) appears as monomer and dimer but was not detected in the high M r fractions. Full-length caspase-2–GFP (Cys mutant) protein expressed in HEK 293T cells was largely insoluble and therefore could not be used in fractionation experiments.