GSM2598158: lef1-/-_3 dpf rep1 (10688X2); Danio rerio; RNA-Seq
Zebrafish: Embryos were fixed for 1.5 hours in 4% PFA/5% sucrose in PBS at RT, followed by whole hypothalamus dissection with super fine forceps (FST, 11252-00). For each biological replicate, 28-38 dissected hypothalami were pooled for lef1 mutant and control samples from at least one single-pair breeding. RNA was extracted using a RecoverAll™ Total Nucleic Acid Isolation Kit for FFPE (Ambion, AM1975) according to the manufacturer's instructions. Three biological replicates were obtained on different days from offspring of different breedings. A total of 300 ng RNA per sample was submitted for RNA-seq. Mouse: E14.5 and P22 non-weaned male Lef1-CON and Lef1-CKO hypothalami were dissected using a fluorescent microscope in ice-cold PBS while tail tissue was retained for genotyping. E14.5 tissues were immediately immersed in RNAlater (Thermo Fisher) and stored at 4 °C. P22 tissues dissected from at least 2 litters were immediately homogenized in TRIzol (Thermo Fisher) and stored at -80 °C. Three biological replicates were prepared from either 5 pooled hypothalami (E14.5) or a single hypothalamus (P22) from Lef1-CON and Lef1-CKO mice, and RNA was extracted on the same day using TRIzol followed by an RNeasy Mini Kit (Qiagen) with on-column DNase digestion (Sigma). 1 µg of RNA per sample was submitted for RNA-seq. Illumina TruSeq Stranded RNA Kit with Ribo-Zero Gold
GEO Accession
GSM2598158