NextSeq 500 paired end sequencing; RNA-Seq of GNRH1 neurons differentiated from human pluripotent stem cells
RNA-Seq of GNRH1 neurons differentiated from human pluripotent stem cells
GnRH enriched cells are GnRH positive cells selected in FACS analysis using our HEL11.4 GNRH1-reporter cell line generated via CRISPR/Cas9.  Cells were sorted and pooled using a FACS machine GNRH1 neurons were differentiated based a protocol previously published (Lund C et al., 2016, Stem cell reports) During the differentiation, after initial treatment with Dorsomporphin and SB431542, FGF8 treatment was continued with DAPT treatment in the terminal step Using RNAqueous micro Total RNA Isolation kit (Thermo Scietific) RNA was isolated. RNA was analyzed for concentration, integrity and quality using Qubit Fluorometer and TapeStation 4200 (Agilent). NEBNext Ultra Directional polyA capture method as paired-end sequencing for read length 75 bp
Experimental Factor: time
20
Experimental Factor: genotype
wild type genotype
Experimental Factor: stimulus
FGF8 100 nanogram per milliliter