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. 2018 Mar 20;12:71. doi: 10.3389/fncel.2018.00071

Figure 5.

Figure 5

Supporting cells (SCs) of the main olfactory epithelium (MOE) take up endocytotic dye after responses to acetylcholine (ACh). (A,B) Confocal images of EEA1 immunolabeling (red). The MOE section was also stained with nuclear marker DAPI (blue in A and cyan in B). The higher magnification fluorescence image overlaid with a transmitted light image in (B) shows strong labeling of EEA1 (red) in supranuclear regions of the SCs. (C–G) Paired images of a SC (left panels) and the same cell showing endocytotic dye pHrodo (red) overlaid with weak light image (right panels): cell in control condition without ACh stimulation (C), cell responsive to 100 μM ACh (D), ACh-nonresponsive cell (E), ACh stimulation in the presence of 0.5 μM atropine (F), and ACh stimulation in a SC isolated from a M3-AChR-KO mouse (G). Scale bars: 10 μm. (H) Average changes in fluorescence levels of the endocytotic dye pHrodo in the experimental conditions used for (A–E). Averaged fluorescence level was measured from an entire cell body. **p < 0.01, ***p < 0.001, one-way ANOVA and Tukey’s post hoc test (n = 5–10).