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The samples were incubated with 100μl reaction mix in a 96 well plate and placed at 37°C

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After fixation, the right ipsilateral sciatic nerve was prepared, and the tissue was immediately incubated in fixation solution for 4 hrs at 4 °C, followed by treatment with 1% osmium tetroxide in 0.1 M PBS for 2 hrs at 4 °C for post-fixation