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. 2019 Oct 11;10:4647. doi: 10.1038/s41467-019-12624-5

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 5 — Sufu Spop mesKO pancreatic mesenchyme shifts towards a gastrointestinal-like signature. a Unsupervised hierarchical clustering analysis of all significantly differentially expressed genes in E13.5 Bapx1^Cre/+;ROSA26^mT/mG;Sufu^f/f;Spop^f/f mutant pancreatic mesenchyme (mutPanc) and control pancreatic (Panc), stomach (St), and intestinal (Int) mesenchyme. Plot is scaled by the Z-score of log-scaled DESeq2 normalized counts, with increasing values (from red to blue) indicating relative enrichment. b Principal component analysis demonstrating that the mutant pancreatic mesenchyme clusters significantly closer to stomach and intestinal mesenchyme as opposed to control pancreatic mesenchyme. c Venn diagram representing common and unique significantly differentially regulated genes between Bapx1^Cre/+;ROSA26^mT/mG;Sufu^f/f;Spop^f/f pancreatic mesenchyme vs. control stomach and intestinal mesenchyme. d, e qPCR analysis of Wnt ligand (d) and Wnt target gene (e) expression in E17.5 Bapx1^Cre/+;ROSA26^mT/mG; Sufu^f/f;Spop^f/f vs. control whole pancreata (n = 3 samples each genotype). f, g Single molecule fluorescent in situ hybridization (smFISH) for Wnt ligand, Wnt2b, in E17.5 control (f) vs. Bapx1^Cre/+;ROSA26^mT/mG;Sufu^f/f;Spop^f/f (g) pancreata co-stained with GFP to mark Bapx1-expressing mesenchyme. h Quantification of average Wnt2b smFISH transcripts per GFP⁺ mesenchymal cell in E17.5 mutants vs. controls (n = 3 samples each genotype). i, j smFISH for Wnt target gene, Axin2, in E17.5 control (i) vs. Bapx1^Cre/+;ROSA26^mT/mG;Sufu^f/f;Spop^f/f (j) pancreata co-stained with GFP to mark Bapx1-expressing mesenchyme. k Quantification of average Axin2 smFISH transcripts per GFP^- epithelial cell in E17.5 mutants vs. controls (n = 3 samples each genotype). White boxes correspond to zoomed-in inserts. Dashed outline denotes border between epithelium and mesenchyme. Data are means ± SEM. n.s. denotes not significant, * denotes p < 0.05, **denotes p < 0.005, *** denotes p < 0.005 by Student’s un-paired, two tailed t-test. Scale bars: 100 µm