Fig. 5. Phorbol ester stimulates the shedding of WT-ACE and GPI-ACE. (A) CHO and (B) SH-SY5Y cells transfected with either WT-ACE or GPI-ACE were incubated for 7 hours in the absence (control) or the presence of either PMA (1 µM), PMA (1 µM) and batimastat (10 µM), carbachol (30 µM), or carbachol (30 µM) and batimastat (10 µM) as indicated. The medium was then harvested and concentrated as described in Materials and Methods. The samples were assayed for ACE activity with BzGly-His-Leu as substrate. Results are the mean ± s.d. of three experiments.