Activation of muscarinic M3 receptors increases constitutive release of sAPPα, but does not affect ADAM10 surface expression. A. HEK-M3 cells were transfected with ADAM10, APP695, or empty vector as indicated. After 48 h cells were pre-incubated for 1 h in serum-free DMEM, then incubated in fresh DMEM in the presence of carbachol (100 μM) or vehicle control (water) for 2 h. Medium was collected, and cells were surface-biotinylated and lysed. Medium extracts were immunoblotted with 6E10 antibodies to sAPPα. Biotinylated proteins were immunoblotted with antibodies to APP or ADAM10. B. Lysates from vehicle-treated cultures were immunoblotted with antibodies to the APP C-terminus. C. Results were quantitated, normalized to control values, and expressed as means ± SEM from 4 or 5 experiments. *Significantly different from vehicle-treated APP695/vector transfectants. **Significantly different from carbachol-treated APP695/vector transfectants.
Carey et al. BMC Cell Biology 2011 12:20 doi:10.1186/1471-2121-12-20