Illumina HiSeq 2000 paired end sequencing; Foxh1/nodal defines context-specific direct maternal Wnt/Beta-catenin target gene regulation in early development
Foxh1/nodal defines context-specific direct maternal Wnt/Beta-catenin target gene regulation in early development
Adult female and male X. tropicalis were primed with 10 units of human chorionic gonadotropin (HCG) 12 hours before boosting. 5 hours before embryo collection, female and male frogs were boosted with 200 units and 100 units of HCG, respectively. For natural mating, single pairs of female and male frogs were placed together, and embryos were harvested approximately every 1 hour after the onset of egg laying. For in vitro fertilisation, testes were dissected from euthanized male frogs and kept in 1x Modified Barth's Saline (MBS) + 0.1% BSA on ice. The testes were subsequently minced and used to fertilise eggs. Fertilised embryos were dejellied in 3% cysteine/0.1x Marc's Modified Ringer (MMR), washed with 0.1x MMR, and kept at 18°C until they developed to desired stages. Xenopus tropicalis embryos were injected into both ventral blastmeres at the four-cell stage with either a beta-catenin morpholino or a beta-catenin morpholino + beta-catenin RNA Total RNA was isolated using the RNeasy Mini Kit (Qiagen) according to manufacturer's instructions. RNA sequencing libraries were prepared using the Illumina TruSeq RNA library (HT) preparation kit
Experimental Factor: developmental stage
mid-blastula stage
Experimental Factor: immunoprecipitate
not applicable
Experimental Factor: protocol
RNA Seq
Experimental Factor: treatment
Beta-catenin Morpholino + RNA reinstatement