The analysis was performed using homogenous cultures of NPCs differentiated from hESC H9, as described in ref. 29. To induce neuronal committed cells (NCCs), NPCs were treated with 20 ng ml−1 BDNF (Peprotech), 20 ng ml−1 GDNF (Peprotech), 1 mM dibutyryl-cyclic AMP (Sigma), and 200 nM ascorbic acid (Sigma)29 for two days. Plasmids and transfections of NPCs are described in Supplemental Methods. NPCs were transfected with control DNA or FGFR1(SP-/NLS)(TK-) and 24 h later were stimulated for 48 h with neuronal differentiation media induced by cAMP/BDNF/GDNF. Experiments were performed using three biological replicates of culture and transfection conditions. The RNAseq protocol was described previously19,29 and is summarized in the Supplementary Methods. The results of RNAseq have been deposited on NCBI GEO with accession code: GSE103307.
