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. Author manuscript; available in PMC: 2014 May 1.
Published in final edited form as: Stem Cell Res. 2013 Feb 5;10(3):428–441. doi: 10.1016/j.scr.2013.01.010

Figure 5. DNA Fragmentation in ETO-induced PCD.

Figure 5

A. ESCs were treated as indicated, fixed, and stained with antibody to caspase-activated DNase (CAD). Nuclei were stained with Draq5. B. Western blot on ESCs treated as indicated and probed for antibody to ICAD or actin. C. ESCs were treated as indicated and probed with antibody to apoptosis-inducing factor (AIF). Draq5 was used to stain nuclei. D. Immunofluorescent staining of EndoG in ESCs treated as indicated. Note that a large portion of EndoG translocates into the nucleus after treatment. E. Western blot showing the level of knockdown of EndoG using two different siRNAs in ESCs after treatment with ETO and harvested at the 4/48 time point. Protein lysates were generated from remaining attached cells. GFP was used to show level of transfection between the transfected cells. F. DNA fragmentation assay on DNA isolated from 4/48 ETO-treated cells, either floating or attached after transfection with control or EndoG siRNAs. G. Flow cytometric analysis of levels of death in ESCs following transfection of EndoG siRNAs and ETO treatment. *p<0.05; #p<0.01