Fig. 4.

Intact cell photolabeling of NPC1 in the presence or absence of NPC2. NPC2-deficient human fibroblast (NPC2 def. hf) cells (8.0 × 10⁶ cells) transiently transfected with pNPC1-YFP only (lane 1) or with pNPC1-YFP and pNPC2-myc-his (lane 2) were photolabeled by using [³H]AC. DMEM without phenol red, instead of Hanks' buffer, was used as the incubation buffer. The samples were processed for radioluminography (RL; exposure time, 7 days) and Western blotting (WB; with rabbit anti-GFP antibodies or with rabbit polyclonal anti-NPC2 antibodies as indicated). Lanes: 1, without pNPC2-mychis; 2, with pNPC2-myc-his. Results are representative of two experiments.