Fig. 5

Mast cells store and release noradrenaline. A Western blotting images of tyrosine hydroxylase (TH) indicate that BMMCs do not express TH, suggesting that BMMCs could not synthesize noradrenaline. B Immunofluorescent images using noradrenaline antibodies in BMMCs treated with DMSO (left) or 10 nM noradrenaline (right) for 24 h. After noradrenaline treatment, the signal was much stronger than that of negative control, suggesting that BMMCs could take up extracellular noradrenaline into the cytoplasm. Scale bars: 20 µm. Four independent experiments were performed, and the representative images are shown. C and D Representative images of immunofluorescent staining in mouse skin (C) and mammary gland (D) using antibodies against Oct3 (green), mMCP-4 (red), and c-kit (white). Note that noradrenaline-storing CTMCs are positive for Oct3. Scale bars: 20 µm. HF hair follicle, Duct mammary duct. Three independent experiments were performed in each organ, and the representative pictures are shown in this Figure. Representative images of negative controls for primary antibodies are shown in Additional file 1. E Stimulation of noradrenaline-storing BMMCs with ionomycin indicates that BMMCs could release noradrenaline. Scale bars: 20 µm. Four independent experiments were performed, and the representative images are shown. F Representative image of immunoelectron microscopic analysis of ionomycin-stimulated BMMCs using noradrenaline antibodies and secondary antibodies labeled with 10 nm gold nanoparticles. Note that the particles are found in the secreting granule (arrow). Scale bars: 1 µm, 200 nm, or 500 nm. Two independent experiments were performed, and a representative image is shown. G Representative images of shRNA-induced BMMCs treated with NeuroSensor 521, confirming that Oct3 is required to take up noradrenaline. Scale bars: 20 µm. Two independent sets of experiment were performed, and the representative images are shown