Retinoic Acid treatment of Xenopus laevis foregut endoderm explants during developmental window of NF15-25

To define Retinoic Acid regulated genes in Xenopus laevis foregut endoderm during developmental window of NF15 to NF25, we treated micro-dissected foregut endoderm explants with 3 doses of Retinoic Acid (25nM, 200nM, 500nM) and performed bulk-RNA seq analysis. Overall design: Foregut endoderm was micro-dissected from wild-type Xenopus laevis embryos at stage NF15 and cultured to stage NF25, a period of approximately 16 hours at 15 degrees C (Adult Xenopus NXR_0031 obtained from National Xenopus Resource Stock Center, Woods Hole, MA; RRID:SCR_013731). Pools of n=8 foregut endoderm explants were harvested at NF25 as 1 biological replicate. 3 such biological replicates per condition were generated (0.1% DMSO treated, 25nM Retinoic Acid (RA, Sigma cat.no. R2625) treated, 200nM RA treated, or 500nM RA treated). RNA was extracted using the Machery-Nagel RNA isolation kit (740955) according to manufacturer's instructions.