TRAP-seq of VMH Nr5a1 neurons

To identify transcripts enriched in ventomedial hypothalamus (VMH) neurons, we used translating affinity ribosome purification combined with RNA-seq (TRAP-seq) from Nr5a1-Cre mice. Nr5a1-Cre mice were crossed to a ROSA26 Cre-dependent EGFP-L10a line. The VMH was microdissected and ribosomes were isolated using an anti-GFP-coated beads. RNA was isolated from the beads and the supernatant and converted to cDNA. Illumina libraries were prepared and sequenced on an Illumina HiSeq machine for 52 cycles.