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. 2012 Aug 2;8(8):e1002842. doi: 10.1371/journal.ppat.1002842

Figure 2. Degradation of BH3-only proteins in Chlamydia-infected cells is prevented by inhibiting CPAF during cell processing.

Figure 2

(A) Lysates of uninfected (0 hpi) or infected HeLa cells were prepared in RIPA buffer (left panel) or by direct lysis in 8M urea (right panel) at the indicated times, separated by SDS-PAGE and analyzed by immunoblotting with antibodies to the proapoptotic BH3-only proteins Puma, Bik, or Bim. Equal loading was monitored for each blot with antibodies to Erk 1/2, but only the loading control for the Puma blot is shown as an example. (B) Uninfected or Chlamydia-infected HeLa cells were treated with 1 µM staurosporine to induce apoptosis, which was monitored by the loss of full-length caspase-3. Immunoblots of the lysates were probed with antibodies to caspase-3, MOMP (marker of Chlamydia infection), or Erk 1/2 (loading control).