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. 2014 Jul 23;33(19):2171–2187. doi: 10.15252/embj.201387244

Figure 4. BID induces mitochondrial release of SMAC in Shigella-infected cells.

Figure 4

  1. HeLa wt cells were left untreated (−) or were infected with Shigella M90T (MOI 30). At the indicated time points whole-cell lysates (WL) and isolated mitochondria (Mito) were analyzed by Western blotting.
  2. HeLa wt cells were transiently transfected with four different siRNAs against BID (siBID1–4) or non-targeting ctrl (siScr). After 48 h cells were analyzed by Western blotting.
  3. HeLa wt cells were transiently transfected with BID-siRNA4. After 48 h cells were left untreated (−) or were infected with Shigella M90T (MOI 30). The mitochondrial fraction was analyzed 6 h p.i. by Western blotting.
  4. HeLa wt cells were transiently transfected with four different siRNAs against BID (siBID1–4) or non-targeting ctrl (siScr). After 48 h cells were left untreated (−) or were infected with Shigella M90T (MOI 30). Cytosolic extracts were analyzed 6 h p.i. by Western blotting.
  5. MEFs isolated from wt or BID−/− mice were left untreated (−) or infected with Shigella M90T (MOI 50). Cytosolic fractions were analyzed by Western blotting at the indicated time points p.i..
  6. HeLa wt cells were transiently transfected with specific siRNAs for BID, NOXA or non-targeting ctrl (siScr). After 48 h cells were infected with Shigella M90T (MOI 30). NF-κB DNA binding activity was analyzed by ELISA at the indicated time points p.i.. Data are presented as mean ± SD (n = 3).
  7. MEFs were treated as in (E). NF-κB DNA binding activity was analyzed by EMSA at the indicated time points p.i..
  8. MEFs were treated as in (E). IL-6 secretion was monitored by ELISA in supernatants of the cells 9 h p.i..

Source data are available online for this figure.