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. 2012 Jan 17;287(10):7766–7779. doi: 10.1074/jbc.M111.290718

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© 2012 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 4. — Caspase-3/ICAD/CAD axis is properly activated in STP-treated SK-N-AS cells. A, protein extracts from SK-N-AS cells treated with 1 μm STP for the indicated times were analyzed by Western blotting to assess caspase-3 activation (upper panel). The membrane was re-probed with a monoclonal antibody to analyze the cleavage of ICAD (middle panel) and subsequently with an antibody against DFF40/CAD (lower panel). The bands corresponding to the unprocessed proteins or the specific fragments are indicated on the right of the panel. Protein loading was assessed by staining the membranes with naphthol blue (NB). B, shown are PCR products of dff40/cad from SK-N-AS cells amplified by either random hexamers (RH) or a specific reverse primer (R) (see “Experimental Procedures”) (upper panel) and cDNA sequencing (lower scheme). The data obtained reveal two silent polymorphisms compared with the nucleotide consensus sequence for human dff40/cad gene (NCBI reference sequence NM_004402.2) at 531 (G to A) and 954 (A to G) nucleotides of the coding sequence (framed in red boxes in the lower scheme). MW, molecular weight marker.