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. 2013 Feb 21;288(13):9200–9215. doi: 10.1074/jbc.M112.411371

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© 2013 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 1. — The apoptotic nuclear collapse and disassembly induced by STP in SK-N-AS cells requires the expression of DFF40/CAD endonuclease. A, SK-N-AS cells were transfected with an NR siRNA or five different CAD siRNAs (1–5). Total protein extracts of the different conditions were obtained, and DFF40/CAD protein levels (upper panel) were analyzed by Western blot analyses 5 days after transfection. The membrane was reprobed with anti-ICAD antibody (lower panel). B, SK-N-AS cells transfected with NR siRNA or CAD siRNA number 2 were treated with 1 μm STP for 6 h or left untreated (Control), and nuclear morphology was analyzed by staining the nuclei with Hoechst 33258. Scale bar = 40 μm. In each condition of treatment, the lower panels are magnifications of the insets in the upper panels. C, electron microscopy from SK-N-AS cells transfected and treated as in B. Scale bars = 40 μm. D, time course analysis of cell death by trypan blue exclusion assay. SK-N-AS cells transfected with NR (white bars) or DFF40/CAD (black bars) siRNA were treated with 1 μm STP.