FIGURE 8.

DC2 and KCP2 interact with the γ-secretase complex. SH-SY5Y cells were transfected with V5-tagged DC2 and KCP2 in a mammalian expression vector as shown (DC2V5 and KCP2V5 panels). Cells were harvested 18 h after transfection and lysed in immunoprecipitation buffer containing 1% CHAPSO, and associated cellular components were then recovered by immunoisolation using antibodies specific for PS1, PEN-2, nicastrin (Nic), APP, SPC25, DC2, or KCP2. The resulting material was analyzed by Western blotting with monoclonal antibody specific for the V5 epitope tag. A fraction of the total cellular extract from the transfected cells (about one-tenth that was used for immunoprecipitation) was included as a control (1/10th vol). IP, immunoprecipitation; WB, Western blot.