ZFIN ID: ZDB-FIG-151120-17
Won et al., 2015 - Lnx2 ubiquitin ligase is essential for exocrine cell differentiation in the early zebrafish pancreas. Proceedings of the National Academy of Sciences of the United States of America   112(40):12426-31 Full text @ Proc. Natl. Acad. Sci. USA
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage Range: Prim-25 to Long-pec
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage: Long-pec

Fig. S2 lnx2a knock-down affects development of the exocrine pancreas. (A) RT-PCR reveals that lnx2a-MO injection efficiently blocks pre-mRNA splicing between exon 2 and exon 3. (B) A schematic drawing indicates the location of primers in exon 2 (F2) and exon 3 (R2). (C and D) lnx2a is involved in the maintenance of ventral pancreas but not initial establishment of endoderm patterning. (C) At 36 hpf, foxa3, gata6, hhex and pdx1-positive cells develop normally in lnx2a-MO injected embryos. (D) At 48 hpf, pdx1-expressing pancreas precursor tissue is normally formed, but mnr2a and ptf1a-postive ventral pancreas precursors are reduced in lnx2a- MO injected embryos (D5 and D6). (E) Detection of lnx2a-MO effect using transgenic embryos and immuno staining. Tg(ptf1a:EGFP) embryos were injected with lnx2a-MO and stained for insulin (Ins) and glucagon (Glu) to visualize β-cells and α-cells, respectively. Confocal images at 48 hpf are shown along three axes. lnx2a-MO injected embryos display defective ventral pancreas (ptf1a: EGFP+), but normal dorsal pancreas-derived β-cells (Ins+) and α-cells (Glu+). (F) Western blot shows the specificity of the anti-Lnx2a polyclonal antibody. Flag-tagged zebrafish Lnx2a, Lnx2b and human LNX2 expression vectors were transfected into 293T cells, and lysates were analyzed by immuno blotting with anti-Lnx2a, LNX2, Flag and Tubulin antibodies, as indicated. Li, Liver; Pa, Pancreas; In, Intestine; DP, Dorsal pancreas; VP, Ventral pancreas; Scale bar, 100 µm (C5) and 10 µm (E1).

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP jh1Tg control Long-pec anterior pancreatic bud IFL
jh1Tg + MO1-lnx2a control Long-pec anterior pancreatic bud IFL
foxa3 AB control Prim-25 endoderm ISH
Prim-25 liver ISH
Prim-25 posterior pancreatic bud ISH
AB + MO1-lnx2a control Prim-25 endoderm ISH
Prim-25 liver ISH
Prim-25 posterior pancreatic bud ISH
gata6 AB control Prim-25 liver ISH
AB + MO1-lnx2a control Prim-25 liver ISH
gcga jh1Tg control Long-pec pancreatic A cell IHC
jh1Tg + MO1-lnx2a control Long-pec pancreatic A cell IHC
hhex AB control Prim-25 liver ISH
Prim-25 posterior pancreatic bud ISH
AB + MO1-lnx2a control Prim-25 liver ISH
Prim-25 posterior pancreatic bud ISH
ins jh1Tg control Long-pec pancreatic B cell IHC
jh1Tg + MO1-lnx2a control Long-pec pancreatic B cell IHC
mnx2a AB control Long-pec anterior pancreatic bud ISH
AB + MO1-lnx2a control Long-pec anterior pancreatic bud ISH
pdx1 AB control Prim-25 pancreatic bud ISH
Long-pec pancreas ISH
Long-pec pancreatic bud ISH
AB + MO1-lnx2a control Prim-25 pancreatic bud ISH
Long-pec pancreas ISH
Long-pec pancreatic bud ISH
ptf1a AB control Long-pec anterior pancreatic bud ISH
AB + MO1-lnx2a control Long-pec anterior pancreatic bud ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
AB + MO1-lnx2a control Long-pec anterior pancreatic bud mnx2a expression decreased distribution, abnormal
Long-pec anterior pancreatic bud ptf1a expression decreased distribution, abnormal
jh1Tg + MO1-lnx2a control Long-pec anterior pancreatic bud EGFP expression decreased distribution, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Proceedings of the National Academy of Sciences of the United States of America for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Proc. Natl. Acad. Sci. USA