Mouse anti beta 3 tubulin

Immunohistochemical (IH) assay was performed as described previously (Lacroix et al., 2014 (link)). Tissue sections were first blocked with 3% goat serum and then incubated with rabbit anti-BAF45D (1:100, Proteintech, Chicago, IL, United States), rabbit-anti-NESTIN (1:100, Sigma, St. Louis, MO, United States), mouse anti-NEUN (1:100, Millipore, Belecula, CA, United States) and mouse anti-beta-III-tubulin (1:200, Arigo) antibodies overnight at 4°C. Positive immunostaining was then detected using a NIKON Eclipse 80i fluorescent microscope and a NIKON Eclipse Ti-S inverted fluorescence microscope. Specific details of the animals and tissues used for IH are given in Supplementary Table S1.

Lysates were prepared from fresh tissues and cells and then subjected to immunoblotting (IB) in accordance with a previously described protocol (Liu et al., 2017 (link)). Proteins were then detected using a range of antibodies: mouse anti-GFAP (1:2000, Proteintech, Chicago, IL, United States), mouse anti-NEUN (1:500, Millipore, Belecula, CA, United States), rabbit anti-BAF45D (1:500, Proteintech, Chicago, IL, United States), mouse anti-beta-III-tubulin (1:1000, Arigo) and rabbit anti-GAPDH (1:2000, Proteintech, Chicago, IL, United States). Specific details of the antibodies used for IF and IB are given in Supplementary Table S2.