SNAP24 and SNAP29 mediate the fusion of secretory carriers with the plasma membrane.

<p>A) Clone 3 cells were mock transfected (TransFast only) or transfected with dsRNA targeting the indicated genes. After 96 hours, the cells were incubated with DD solubiliser at 25°C for 80 minutes and their mean fluorescence determined using flow cytometry. The amount of cargo remaining in the cells was calculated and plotted (Error bars show experimental range for three repeats). B) Clone 3 cells were mock transfected (TransFast only) or transfected with dsRNA targeting the indicated genes. After 96 hours, the cells were incubated with DD solubiliser at 25°C for 80 minutes and the media and cells harvested for immunoblotting. Solid arrowhead indicates unprocessed cargo and unfilled arrowhead furin processed cargo. The amount of processed GH in the media was quantified using densitometry and plotted in C (Error bars show experimental range for two repeats). D) Clone 3 cells were either mock transfected or transfected with dsRNA targeting the indicated genes. After 72 hours the cells were seeded onto coverslips. The next day, the cells were incubated with AP21998 at 25°C for 80 minutes. The cells were then fixed and stained for the Golgi marker GM130. Scale Bar 10μm.