br Materials and methods br Acknowledgements This study

Materials and methods
Acknowledgements This study was supported by the European Union within the 7th European Community Framework Programme through funding for the NEUROMICS network (F5-2012-305121 to L.S.), the Marie Curie International Outgoing Fellowship (grant PIOF-GA-2012-326681 to R.S. and L.S.), and the DZNE intersite project (grant to L.S.). We thank the patient for participation.
Resource table. Resource details Peripheral blood was collected from a 39-year-old female donor, a mother of two sons with genetically characterized MPS II disorder diagnosed by Department of Pediatrics, University of Pécs (Hungary). Both children carry a pathogenic X-linked triptolide of the IDS gene (NM_000202.7(IDS):c.85C>T, p. Gln29Ter). The single nucleotide variation (SNV) results a premature termination codon, leading to Iduronate 2-sulfatase enzyme deficiency and to the severe accumulation of glycosaminoglycans (Wraith et al., 2008). MPS II is an X-linked recessive disorder which manifests almost exclusively in males, while females are mainly unaffected carriers with a mutant and a wild-type allele (Neufeld et al., 1977; Pinto et al., 2010). The presence of both the mutant and wild-type allele was first confirmed in the donor PBMCs (data not shown), then in the MPSII-mother.1 iPSC line by Sanger sequencing of the PCR product harboring the pathogenic SNV (Table 1, Fig. 1A). To generate the MPSII-mother.1 iPSC line the pRRL.PPT.SF.hOKSMco.idTomato.preFRT lentiviral vector was used, which was shown that under certain condition is self-silencing shortly after transduction (Voelkel et al., 2010; Warlich et al., 2011). TRA-1-60 expressing iPSC-like colonies were picked 18–21days post-transduction based on in vivo immunocytochemistry staining (ICC) (Fig. 1B). Six stable lines were maintained and based on morphological criteria the MPSII-mother.1 was chosen for further examination. The pluripotency of MPSII-mother.1 line was confirmed by alkaline phosphatase staining (ALP) and by ICC for endogenous NANOG and E-CADHERIN (Fig. 1B) after silencing of the exogenous hOKSM construct. The in vitro spontaneous differentiation potential of the iPSC line towards the three germ layers was demonstrated by the expression of ectodermal (βIII-TUBULIN), mesodermal (BRACHYURY) and endodermal (GATA4) markers (Fig. 1B) using ICC. The karyotype of the MPSII-mother.1 iPSC line was determined by Giemsa-banding, proving normal diploid 46, XX karyotype, without any detectable abnormalities (Fig. 1B).
Materials and methods
Acknowledgement We would like to thank for the Department of Pediatrics, University of Pécs (Hungary) for collecting blood samples of donors and providing clinical data. This work was supported by grants from EU FP7 projects (IDPBYNMR, PITN-GA-2010-264257; Anistem, PIAPP-GA-2011-286264; EpiHealth, HEALTH-2012-F2-278418); and Research Centre of Excellence – 11476-3/2016/FEKUT.
Resource table. Resource details The generation of human IPSC line i10984 was carried out using E8 based media and feeder-free conditions. EBV-transformed B-lymphocyte cells from an 83-year old Alzheimer\'s Disease (AD) patient, generated by Coriell Institute For Medical Research, were employed. The patient had a 15year-history of progressive presenile dementia with a parent and three siblings of the patient also being affected (iPSC line i11001). The patient was homozygous for the APOE4 allele. This was confirmed by a PCR-RFLP assay (Hixson and Vernier, 1990) carried out with the lymphoblastoid cell line. (Fig. 1A). The lymphoblastoid cells (LCs) were electroporated with OriP/EBNA-1 based episomal plasmids expressing reprogramming factors OCT4, SOX2, KLF-4, L-MYC, LIN28 and a p53 shRNA (Okita et al., 2011). The APOE4/4 genotype of the generated iPS line was confirmed by sequencing (Fig. 1A). After several passages, a PCR for episomal vector backbone confirmed that exogenous episomal factors were not being expressed anymore (Fig. 1A).