In situ hybridization was performed on fresh frozen sagittal brain sections (18 μm), using digoxigenin (DIG) -labeled antisense probes for murine Plp1/Dm-20, Mbp and Enpp6 as described previously30,53,54. The probe was synthesized by T3 RNA polymerase (Promega, Cat#P208C) and labeled with DIG RNA label mix (Roche, Cat#11277073910). An anti-DIG antibody conjugated with alkaline phosphatase (Roche, Cat#11093274910) was used to probe sections and followed by stained with 5-bromo-4-cloro-3-indolyl phosphate (BCIP) (Solarbio, Cat#B8090) /nitro blue tetrazolium (NBT) (Solarbio, Cat#N8140) chromogenic substrates.
