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Primary antibodies against β(III)tubulin (1:400, rabbit anti-β(III)tubulin monoclonal antibody [EP1569Y]-Alexa Fluor ® 647, #ab190575, Abcam, United Kingdom) and O4 (1:400, mouse anti-O4 IgM, #MAB1326, R&D systems, United States) were incubated overnight in PBS containing 0.01% Triton-X and 2% GS at 4°C
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