Figure 6: Effects of hBD-3 on the expression of truncated Bid and Bcl-xL. Neutrophils (106 cells/mL) were incubated at 37°C for 4 h in RPMIFBS in the absence (Control) or presence of hBD-3 (5 μg/mL). Expression of truncated Bid (tBid) and Bcl-xL was detected by probing with goat anti-human/mouse Bid polyclonal Ab (a) or mouse anti-Bcl-xL mAb (b), and HRP-conjugated rabbit anti-goat IgG or HRP-conjugated goat anti-mouse IgG/IgM, respectively. To confirm that equal amounts of proteins were analyzed in each sample, the blots were stripped, and GAPDH was detected by reprobing with mouse anti-GAPDH mAb and HRP-conjugated goat anti-mouse IgG/IgM. Anti-human/mouse Bid polyclonal Ab can recognize 17-kDa truncated Bid (tBid) as well as 22-kDa Bid (upper half of panel (a)). Data are from 1 of 4 separate experiments.