![Figure 2. O2− inhibits PP2A-mediated dephosphorylation of Bcl-2 at S70. (A-C) Lysates from DDC-treated Jurkat, Hela, and MDA231 were immunoprecipitated (IP) and immunoblotted (IB) using the indicated antibodies. Input refers to whole cell lysates that are not subjected to IP procedures. Immunoglobulin G (IgG) refers to untreated lysates that were incubated with nonspecific IgG primary antibodies. (D) Mitochondria isolated from MDA231 or Jurkat cells were incubated with proteinase K for 25 minutes followed by lysis and IB for the indicated proteins. As a control, incubation of mitochondria with Triton-X100 resulted in the digestion of SOD2 (matrix protein) and COX Va (inner mitochondrial membrane [IMM] protein). (E) Mitochondrial/cytosolic fractions from DDC-treated Jurkat or MDA231 cells (with or without tiron pretreatment) were IB for the indicated proteins. SOD1 was employed as a cytosolic marker, whereas prohibitin, an IMM protein, served as a mitochondrial marker. (F) Confocal fluorescence imaging studies of DDC-treated MDA231 cells. Colocalization of Bcl-2 (red) and PP2A-C (green) appears as yellow punctate (red arrows). (G) Confocal imaging of Jurkat cells 48 hours after silencing of SOD1. Selected cells were enlarged for greater clarity.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/124/14/10.1182_blood-2014-03-563296/4/2223f2.jpeg?Expires=1735497658&Signature=jTq43AXAUFgBCs8Yy2pdt30sTFoxHlhfuXrzJ3ae9-SMTkkwoQL4Sm3rxPjRMfHrUrOx1PiSsancVAkCsU6cFaXbknUlvBirS~xXUOLKHqxZTNJnIRRQZVqrjM~WwQBpog5RrwQxyp6qTlpM80Hw5RcWCxyjg4Hc~DXQ7vaXhjWnSTK4mAkss0ar7wYss6at5Z6b7XWJxO6eWSbmH1hoZKHVrLNidvW0NkGmBGUGpOraV709vk1OlFQx3Piub6b7W8aSVFIpvndgA0~qWX3Xm~NvRCxvkLd0eyGqEO-46gG4O9XhmjWgMv6wEEZje-YH3H5YlFp9q3tITuhTM5FIgA__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
O2− inhibits PP2A-mediated dephosphorylation of Bcl-2 at S70. (A-C) Lysates from DDC-treated Jurkat, Hela, and MDA231 were immunoprecipitated (IP) and immunoblotted (IB) using the indicated antibodies. Input refers to whole cell lysates that are not subjected to IP procedures. Immunoglobulin G (IgG) refers to untreated lysates that were incubated with nonspecific IgG primary antibodies. (D) Mitochondria isolated from MDA231 or Jurkat cells were incubated with proteinase K for 25 minutes followed by lysis and IB for the indicated proteins. As a control, incubation of mitochondria with Triton-X100 resulted in the digestion of SOD2 (matrix protein) and COX Va (inner mitochondrial membrane [IMM] protein). (E) Mitochondrial/cytosolic fractions from DDC-treated Jurkat or MDA231 cells (with or without tiron pretreatment) were IB for the indicated proteins. SOD1 was employed as a cytosolic marker, whereas prohibitin, an IMM protein, served as a mitochondrial marker. (F) Confocal fluorescence imaging studies of DDC-treated MDA231 cells. Colocalization of Bcl-2 (red) and PP2A-C (green) appears as yellow punctate (red arrows). (G) Confocal imaging of Jurkat cells 48 hours after silencing of SOD1. Selected cells were enlarged for greater clarity.
