Pd 0332991 isethionate

Dorsomorphin dihydrochloride, SB431542, CHIR99021 and Purmorphamine were purchased from Stemgent (Cambridge, MA, USA). Y27632, PD 0332991 isethionate and SU9516 were purchased from Tocris (Bristol, UK). Recombinant human GDNF, BDNF, NGF, bFGF and TGF-β3 were purchased from PerproTech (Rocky Hill, NJ, USA). cAMP, Ara-C, Ara-A, ascorbic acid and N-acetyl-L-cysteine were purchased from Sigma (St. Louis, MO, USA). FUW-tetO-LoxP-hOCT4, shP53pLKO.1, pMD2.G and psPAX2 were purchased from Addgene (Cambridge, MA, USA). Human Ascl1 (Genebank accession BC031299), Foxa2 (BC011780), Lmx1b (BC113491), FEV (BC023511.2), Nurr1 (CV028069), Lmx1a (BC06635), NeuroD1 (NM_002500), NeuroD2 (NM_006160) and Pitx3(NM_005029) were purchased from Open Biosystems (Pittsburgh, PA, USA) and subcloned by PCR to the EcoRI site on the FUW-tetO-LoxP vector. Human miR-124 (MIMAT0000422) and Nkx2.2 (NM_002509) were amplified from normal human fibroblast genomic DNA and subcloned to the EcoRI site on the FUW-tetO-LoxP vector. FUW-LoxP-M2rtTA was generated by subcloning the BspEI fragment containing the loxP site from FUW-tetO-LoxP-hOCT4 (Addgene) to the BspEI site on the 3′LTR of FUW-M2rtTA (Addgene). All constructs were verified by sequencing.