Figure 4

From: Caspase-mediated proteolysis of the sorting nexin 2 disrupts retromer assembly and potentiates Met/hepatocyte growth factor receptor signaling

Figure 4

SNX2 is a caspase-6 substrate. (a) HeLa cells were transfected with control (siCTL) or caspase-6 small interfering RNAs (siRNAs) (siC6) followed by a 16 h treatment with TRAIL (100 ng/ml), STS (0.5 μM) or UV (254 nm wavelength; 100 J/m2). (b) Eighty micrograms of HeLa cell extracts were left untreated or incubated with 250 nM active site-titrated recombinant caspase-6 for an hour at 37 °C. (c) HEK293T cells were transfected with either control (−), Flag-tagged WT or catalytic mutant (C285A) caspase-6 plasmids for 48 h. Proteins were analyzed by immunoblotting using the indicated antibodies. Closed and open arrowheads indicate full-length and cleaved fragments, respectively. Actin and HSP90 were used as loading controls. The asterisk indicates a nonspecific protein recognized by the SNX2 antibody.