Figure 4.

TNF-α/CHX-induced apoptosis was dramatic in Prdx6-overexpressing Prdx6^KD cells. (a) Prdx6 was transfected into Prdx6^KD cells. The expression of Prdx6 was compared in wt BEAS-2B, Prdx6^KD, and Prdx6-overexpressing Prdx6^KD cells with an antibody specific for Prdx6. (b) The wt BEAS-2B and Prdx6-overexpressing Prdx6^KD cells were treated with TNF-α/CHX for various times as indicated. The lysates were examined by western blotting using anti-caspase-3, anti-caspase-8, and anti-GAPDH. (c and d) The wt BEAS-2B and Prdx6-overexpressed Prdx6^KD cells were treated with TNF-α/CHX for different times as indicated. Caspase-3 (c) and caspase-8 (d) activities were measured using the CaspACE kit according to the manufacturer's instructions. The results are expressed as mean±S.D. for triplicate assays. (e) The wt BEAS-2B and Prdx6-overexpressed Prdx6^KD cells were treated with TNF-α/CHX for various times as indicated. The cells were stained for AnnexinV. The percentage of AnnexinV-positive cells was analyzed with the FACSCalibur system and determined with the CellQuest software. The results are expressed as mean±S.D. for triplicate assays