BAX/BAK Independent Cell Death at: doi:10.1371/journal.pone.0005646.s005 (1.75 MB Bax/Bak Null MEFs+Bid BH3peptide Mitotracker at: doi:10.1371/journal.pone.0005646.s006 (0.63 MB tagged to FITC for 5 minutes. Flow cytometry was performed to assess for GFP positive cells. Found at: doi:10.1371/journal.pone.0005646.s001 (0.53 MB TIF) Figure S2 Bax2/2/Bak2/2 MEFs overexpressing BCL-XL are not protected against peptide induce cell death. (A) LDH release of Bax2/2/Bak2/2 MEFs overexpressing empty construct or BCL-XL when treated with increasing concentration of BID or mutant BID BH3 peptide at 24 hours. Mean values6SEMs of 3 independent experiments are shown. (B) Western Blot analysis of whole cell lysate of Bax2/2/Bak2/2 MEFs, overexpressing empty construct or BCL-XL. Found at: doi:10.1371/journal.pone.0005646.s002 (0.69 MB TIF) Figure S3 CsA inhibits the permeability transition pore of Bax2/2/Bak2/2 MEFs. Bax2/2/Bak2/2 MEFs were loaded with Calcein AM in the presence of cobolt choride for mitochondria labeling. Following mitochondrial Calcein uptake, only cells that were treated with CsA allowed for Calcein retention following treatment with ionomycin. Found at: doi:10.1371/journal.pone.0005646.s003 (0.50 MB TIF) Movie S1 Bax/Bak Null MEFs Mitotracker Red Found at: doi:10.1371/journal.pone.0005646.s004 (0.85 MPG) MB Movie S2 Wild-type MEFs Mitotracker Red References 1. Green DR, Kroemer G (2004) The pathophysiology of mitochondrial cell death. Science 305: 626–629. 2. Youle RJ, Strasser A (2008) The BCL-2 protein family: opposing activities that mediate cell death. Nat Rev Mol Cell Biol 9: 47–59. 3. Wei MC, Lindsten T, Mootha VK, Weiler S, Gross A, et al. (2000) tBID, a membrane-targeted death ligand, oligomerizes BAK to release cytochrome c. Genes Dev 14: 2060–2071. 4. Youle RJ, Karbowski M (2005) Mitochondrial fission in apoptosis. Nat Rev Mol Cell Biol 6: 657–663. 5. 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