her6 and her9 are expressed in the same subpopulation of cranial neural crest cells as jag1b, and their expression is dependent upon jag1b function. (A) UMAPs from single cell RNA-sequencing on 24 hpf sorted cranial neural crest cells. As in our previous work (29), cranial neural crest cells can be clustered into four distinct populations. Based on our previous study and the landmark genes enriched in each population we can identify where the cells reside in the intact animal. Arrows indicate jag1b enrichment in a subset of anterior arch cells. Although her6 is broader, arrows indicate that some of the strongest expressing cells are the jag1b enriched populations. her9 is also broadly expressed among cranial neural crest cells, but at lower levels than her6. Arrows indicate her9 enrichment in the jag1b;her6 expressing populations. (B) We performed qPCR on 28 and 48 hpf genotyped heads from wild type and jag1b homozygous mutant siblings. Expression of each her gene was normalized to rps18, then normalized to the relevant wild-type control. Reactions were performed in technical triplicate and the results represent three to six biological replicates. Each biological replicate was pooled heads from 5–6 genotyped homozygous wild types or homozygous mutants. Error bars are standard deviation. Asterisks indicate significant differences from wild type controls at p < 0.05 by students t-test.
