Role of barhl1 in amphbian cerebellum

RNA- sequencing analysis was performed on dissected Rhombomere 1 from Xenopus Laevis tadpoles depleted or not for BarH-like 1 (Barhl1), a direct target gene of Atonal 1 (Atoh1), a master regulator of cerebellar glutamatergic neurons development, and an inhibitor of T-Cell factor activity. X. laevis embryos were injected with three different conditions: MObarhl1-1; MObarhl1-2 and MOct in the two dorsal blastomeres at four cells stage. At stage 42, neural tubes were extracted in RNAse-free conditions, and the rhombomere 1 which includes the URL was carefully dissected. For each condition, three biological replicates were collected. Each replicate contains three rhombomeres. Poly A RNA was purified. Sequencing was performed using Illumina NovaSeq (paired-end sequencing) by Next Generation Sequencing Platform (NGS) (Institut Curie, Paris FR). This transcriptomic analysis identifies Barhl1 cerebellar target genes, and confirms that Barhl1 acts as a gate keeper for progenitor exit from the URL, through silencing of T-Cell Factor transcriptional activity.