For electron microscopy, mice were anesthetized with ketamine/xylazine and perfused with 2% paraformaldehyde and 0.2% glutaraldehyde in phosphate-buffered saline (PBS) (0.1 M, pH 7.4), and dissected sciatic nerves were fixed with 2% paraformaldehyde and 2.5% glutaraldehyde in PBS (0.1 M, pH 7.4) for 1 hour at room temperature and post-fixed in 1% osmium tetroxide in cacodylate buffer (0.1 M, pH 7.2) for 1 hour at 4°C. After washing, tissues were stained in 0.5% uranile acetate (pH 4) for 1 hour at 4°C, then dehydrated through graded alcohols, and embedded in Epon. Semithin sections (500 nm) were stained with toluidine blue before being observed with an optical microscope. Ultrathin sections (50 nm) were analyzed with a transmission electron microscope (JEOL 1200EX, Grenoble Institute of Neurosciences, France) at 80 kV, and images were acquired using a digital camera (Veleta, Olympus). Morphometric measurements were done with ITEM software (Soft Imaging System, Olympus). The same protocol was applied for electron microscopy on JoMa1.3 cells.

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