Expression of BH3-only proteins in DC subsets. (A) T cells were generated by activation of mouse spleen cells with ConA and IL-2. Western blot analyses were performed to detect BH3-only proteins in T cells, or bone marrow–derived DCs with or without treatments with LPS and CpG. (B) RNA was prepared from bone marrow–derived DCs with or without stimulation of CD40 or TLRs for 20 hours and used for semiquantitative RT-PCR for Bim or actin. The 499-bp, 331-bp, and 241-bp bands correspond to BimEL, BimL, and BimS, respectively.28 (C) Cell lysates prepared as in panel A were used for Western blot analyses of BAD and phosphorylated BAD. (D) Cell lysates prepared as in panel A were used for Western blot analyses of antiapoptotic molecules. (E) Freshly purified T cells, mDCs, and pDCs from mouse spleens were lysed for Western blot analysis of Bim and Bid. (F) mDCs and pDCs purified from mouse spleens were treated with anti-CD40 or different TLR stimuli for 24 hours. Cells were lysed for Western blot analyses of Bim and Bid.
