Vascular GM-CSF released by TLR2/6-dependent MALP-2 stimulation occurs exclusively from the endothelium. (A) GM-CSF levels in supernatants of different vascular cells after MALP-2 (1 μg/mL) stimulation for 24 hours determined by ELISA (n = 3-6). SMC indicates smooth muscle cells. **P < .01 vs control. (B) Picture represents a segment of the murine aorta (Zeiss Axiovert 200M microscope, 5×/0.25 NA dry objective, Zeiss AxioCam MRc camera, AxiVision Version 4 software). (C) GM-CSF levels in the supernatants of murine aortic segments after MALP-2 (1 μg/mL) stimulation for 24 hours determined by ELISA. Unspecific IgG (50 μg/mL) was used as antibody control (n = 4-6). AB indicates antibody (25 μg/mL each, ). **P < .01 vs control. ##P < .01 vs MALP-2. (D) Immunofluorescent staining of murine aortic segments with antibodies against CD31. Representative pictures of segments with intact or removed endothelium are shown (Leica DM 4000B microscope, 20×/0.50 NA dry objective, Leica DFC 320 camera, Leica QWin Version 3 software). (E) GM-CSF levels in supernatants of murine aortic segments with intact or removed endothelium after MALP-2 (1 μg/mL) stimulation for 24 hours determined by ELISA (n = 4). **P < .01 vs control. ##P < .01 vs MALP-2. (A,C,E) Error bars represent mean ± SEM.