ZFIN ID: ZDB-FIG-161027-6
Huang et al., 2016 - Sox9b is a mediator of retinoic acid signaling restricting endocrine progenitor differentiation. Developmental Biology   418(1):28-39 Full text @ Dev. Biol.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibody:
Fish:
Conditions:
Anatomical Terms:
Stage Range: Day 5 to Days 7-13
PHENOTYPE:
Fish:
Conditions:
Observed In:
Stage Range: Day 5 to Days 7-13

Fig. 6

sox9b haploinsufficiency impedes the effects of RA on CAC differentiation. (A) Immunofluorescence staining using a Sox9b antibody in 5 dpf pancreata expressing Tp1:nuc-mCherry. Sox9b expression co-localizes with nuclear mCherry in CACs (white arrows). Pancreas is outlined in dashed line. Scale bar=20 µm. (B) Western blot demonstrating that Sox9b expression is reduced in sox9bfh313 heterozygous larvae. (C-F) Images of dissected pancreata from 5 dpf larvae with the following genotype: (C-D) wildtype (sox9b+/+) (E-F) sox9bfh313 heterozygous (sox9b+/). in situ hybridization for neuroD was used to detect 2° islets. Larvae of differing genotypes (as indicated) were treated from 3 to 5 dpf with either DAPT (C, E) alone or RA+DAPT (D, F). 2° islets are indicated by black arrow heads. White dashes outline the pancreata. Scale bar=100 µm. (G) Numbers of CACs quantified using expression of Tp1:eGFP in pancreata from 5 dpf larvae, genotyped as indicated. Heterozygotes do not have significantly different numbers of CACs compared to wildtype. (H) Numbers of 2° islets/pancreas from genotyped larvae after being treated with DAPT alone or RA+DAPT. Error bar represents SD; **p<0.001.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
mCherry jh32Tg standard conditions Day 5 pancreatic centroacinar cell nucleus IHC
neurod1 WT chemical treatment by environment: DAPT Day 5 endocrine pancreas ISH
Day 5 secondary islet ISH
WT chemical treatment by environment: all-trans-retinoic acid, chemical treatment by environment: DAPT Day 5 endocrine pancreas ISH
Day 5 secondary islet ISH
sox9bfh313/+ chemical treatment by environment: DAPT Day 5 endocrine pancreas ISH
Day 5 secondary islet ISH
sox9bfh313/+ chemical treatment by environment: all-trans-retinoic acid, chemical treatment by environment: DAPT Day 5 endocrine pancreas ISH
Day 5 secondary islet ISH
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab2-sox9b IHC jh32Tg standard conditions Day 5 pancreatic centroacinar cell nucleus
WB WT standard conditions Days 7-13 whole organism
WB sox9bfh313/+ standard conditions Days 7-13 whole organism
Phenotype Details
Fish Conditions Stage Phenotype
WT chemical treatment by environment: DAPT Day 5 secondary islet increased amount, abnormal
WT chemical treatment by environment: all-trans-retinoic acid, chemical treatment by environment: DAPT Day 5 secondary islet amount, ameliorated
sox9bfh313/+ standard conditions Days 7-13 whole organism Ab2-sox9b labeling decreased amount, abnormal
sox9bfh313/+ chemical treatment by environment: DAPT Day 5 secondary islet increased amount, abnormal
sox9bfh313/+ chemical treatment by environment: all-trans-retinoic acid, chemical treatment by environment: DAPT Day 5 secondary islet increased amount, abnormal
Acknowledgments:
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Reprinted from Developmental Biology, 418(1), Huang, W., Beer, R.L., Delaspre, F., Wang, G., Edelman, H.E., Park, H., Azuma, M., Parsons, M.J., Sox9b is a mediator of retinoic acid signaling restricting endocrine progenitor differentiation, 28-39, Copyright (2016) with permission from Elsevier. Full text @ Dev. Biol.