VWF-proteins are secreted and biologically active. A: Dose-response curve of NDP-MSH and forskolin showing their ability to stimulate 293T cells transfected with MC4R and cAMP sensitive reporter gene. B: similar as 2A, but in the presence of 10% of conditioned medium from 293T cells transfected with either AAV-GFP, AAV-flAgRP or AAV-VWF-AgRP83-132 . C: similar as 2B, but with 50% of conditioned medium. D: dose-response curve of NDP-MSH and forskolin as positive control for MC4R and CRE-LacZ transfection. E: response curve of cells transfected with MC4R and CRE-LacZ incubated with different dilutions of conditioned medium. Medium was taken from cells transfected with AAV-GFP, AAV-MSH1 or AAV-MSH4. Graphs represent an average of 4 trials.