Map2 hm 2

The primary antibodies for immunostaining were GT335 (Gift from B. Edde and Dr. C. Janke, 1:2000), Neurofilament M (RMO14.9; Cell Signaling Technology, 1:25), and Map2 (ab5622; Merck Millipore, 1:1000). The secondary antibodies were Alexa fluor 488 (Thermo Fisher Scientific, 1:1000) and Alexa fluor 568 (Thermo Fisher Scientific, 1:1000). The antibodies used for immunoblot were TTLL1 (Ikegami et al., 2010, 1:1000), TTLL7 (Ikegami et al., 2006, 1:1000), GT335 (1:20,000), α-tubulin (DM1A; Sigma-Aldrich, 1:50,000), β-tubulin (TUB2.1; Sigma-Aldrich, 1:2000), tyrosinated tubulin (1A2; Sigma-Aldrich, 1:2000), acetylated tubulin (6-11B-1; Sigma-Aldrich, 1:20,000), neurofilament 200 (NE14; Sigma-Aldrich, 1:500), neurofilament 160 (NN18; Sigma-Aldrich, 1:250), glyceraldehyde-3-phosphate dehydrogenase (GAPDH, 6C5; Sigma-Aldrich, 1:5000), CLIP170 (Sigma-Aldrich, 1:500), Map1A (HM-1; Sigma-Aldrich, 1:1000), Map1B (Sigma-Aldrich), 1:1000; Map2 (HM-2; Sigma-Aldrich), 1:1000; Tau-1 (MAB3420; Merck Millipore, 1:10,000), dynein (74.1; Merck Millipore, 1:1000), KIF1A (no. 16; BD Biosciences, 1:500), KIF5 (H2; Merck Millipore, 1:500), and KIF17 (Sigma-Aldrich, 1:1000). The antibodies used for 2D electrophoresis were GT335 (1:20,000), α-tubulin (DM1A; 1:4000), β-tubulin (TUB2.1; 1:4000).

Immunohistochemistry was performed with neuronal cortical cultures of B6.2D2 mice. Neurons were stained for ICAM-5 (TubIII, Covance), NeuN (Millipore, Billerica), MAP2 (HM-2, Sigma), and Tuj1 (Covance); cell nuclei were stained with DAPI (Invitrogen). Pictures were obtained using a confocal laser scanning microscope (Leica TCS-SP8; Leica Microsystems Heidelberg, Mannheim, Germany).