1st EXPERIMENTAL APPROACHES FROM THE PAPER THAT SUPPORTS A ROLE FOR TGF-B/BMP signaling IN THIS DEVELOPMENTAL EVENT:
- First, semi quantitative reverse transcription PCR (qPCR): used to look for a regulator of SNW1 and BMP. Results in Xenopus embryos show SNW1 transcripts present in both dorsal and ventral sides, but at Neurula stages is present more DORSALLY.
- Western blot: showed SNW1 protein levels lowest at earlier stages and continued to increase through the developmental stages.
- Knocking down SNW1: show reduced SNW1 protein levels starting at the level where it was seen to be lowest to start, but NOT before, indicating that the protein prior to this stage is maternally deposited, and the rest is zygotically produced.
- Experiments were repeated in zebrafish, and similar patterns observed.
- SNW1 knockdown: INHIBITS neural crest induction as measured by loss of neural crest markers Snail, Slug and Sox 9 at the injected side at stage 14.