TRAP-seq of VMH LeprnSlc17a6 neurons

To identify transcripts enriched in ventomedial hypothalamus (VMH) neurons that contain both Lepr and Slc17a6, we used translating affinity ribosome purification combined with RNA-seq (TRAP-seq) from Lepr-Cre;Slc17a6-Flpo mice. Overall design: Lepr-Cre;Slc17a6-Flpo mice were crossed to a ROSA26 Cre- and Flp-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.