Fig. 6

Polymicrobial infection activates BAD pro-apoptotic activity in septic shock. Bad^−/− mice and their WT littermates (a, b, e), or Tnf-R1^−/− mice and their WT littermates (c–e) were subjected to high-grade CLP surgery or Sham procedure, as described in Fig. 5a. a–d Total liver tissue extracts were fractionated to determine BAD translocation from the cytosol [(containing cytoskeleton, Cytosol(+C)] to mitochondria (see “Materials and methods” for details) (a, c and Supplementary information, Figure S9n, S9p), or BAD release from the cytoskeleton to the cytosol [containing mitochondria; Cytosol(+M)] (see “Materials and methods” for details) (b, d and Supplementary information, Figure S9o, S9q). The levels of BAD and the percentage of IKK-phosphorylated BAD in total BAD were determined and quantitated, as described in Fig. 1f (See “Materials and methods” for details). e Phosphorylation of IKKβ, IκBα, phospho-BAD(S26), as well as expression levels of IKKβ, IκBα, BAD, TNF-R1, and β-actin in liver were analyzed by immunoblotting with corresponding antibodies. All the results represent three individual experiments with similar results