Loss of function assays were performed after bilateral injections with their respective controls. Wnt knockdown was performed by the combined inhibition of Wnt 1 and 4 by morpholinos. For RNA expression levels, paired single neural folds were finely dissected and total RNA isolated with the RNAqueous-Micro Total RNA Isolation Kit (Thermo Fisher Scientific, #AM1931) according to the kit's protocol. RNA was poly(A) selected using the NEBNext Poly(A) mRNA Magnetic Isolation Module (New England Biolabs, E7490). TruSeq-barcoded RNAseq libraries were generated with the NEBNext Ultra II Directional RNA Library Prep Kit (New England Biolabs, E7760) and sequenced on an Illumina NextSeq500 instrument.
GEO Accession
GSM4520625