Analysis of Xenopus tropicalis stage NF33 regional gene expression in micro-dissected domains along anterior-posterior axis and Hedgehog, Retinoic Acid, and Rfx6 dependent gene expression in micro-dissected NF33 foregut tissue (mesoderm + endoderm)

To define regional gene expression domains at stage NF33 along the Xenopus tropicalis embryonic anterior-posterior axis and also identify Rfx6, Retinoic Acid, and Hedgehog regulated genes at stage NF33. Overall design: Retinoic Acid, and Hedgehog regulated genes at stage NF33. Tissue was micro-dissected from wild-type Xenopus tropicalis embryos at stage NF33 (Adult Xenopus NXR_1018, Xtr.NigerianSt549, obtained from National Xenopus Resource Stock Center, Woods Hole, MA; RRID:SCR_013731). Pools of n=8 explants (mesoderm+endoderm)s were harvested as 1 biological replicate. 3 such biological replicates per condition were generated. RNA was extracted using the Machery-Nagel RNA isolation kit (740955) according to manufacturer's instructions. 40uM cyclopamine (Tocris #1623), 100nM BMS493 (Tocris #3509) treatment from NF15-33. 1ng total Rfx6 morpholino oligo or 5bp mismatch oligo (500pg per cell) was injected at 2-cell stage.