Figure 11.

Defects in peptide secretion are rescued upon tranfection-mediated reintroduction of the wild-type and F115/E132 mutant, of Munc18-1. (A and B) Secretion of NPY-hPLAP from the double knockdown clones (D7 in A; D16 in B) that were transfected with the control plasmid or the Munc18-1 expression plasmid was stimulated by 70 mM KCl for 25 min. Error bar indicates SEM (n = 15 for D7; n = 18 for D16). (C) The expression of transfected Munc18-1 proteins in D7 and D16 clones. Thirty micrograms of total homogenates from D7 and D16 clones that were transiently transfected with Munc18-1 expressing plasmids was analyzed by SDS-polyacrylamide gel electrophoresis and immunoblotting using anti-Munc18-1 and anti-VCP (loading control) antibodies. Signals were detected with enhanced chemiluminescence detection system. A number on the left indicates the position of a molecular weight marker.