Fig. 2. PDGFRahigh cells-assisted Wnt-signalling gradient is present in E16.5 SI and required for the establishment of region-specific organoid cultures.
a RNA-seq of PDGFRahigh cells showing differentially expressed ligands between proximal, mid and distal regions. Highlighted in red are Wnt-pathway components. b RNA-seq results for Axin2 and Lgr5 from sorted EpCAM cells isolated from proximal, mid and distal regions. The y-axis shows RPKM (reads per kilobase per million mapped reads). c Detection of Top.CFP (white), E-cadherin (red) and DAPI (blue) in proximal, mid and distal regions. Scale bars, 20 μm. d–f Formation of organoids from DAPI−/CD31−/CD45−/EpCAM+ cells isolated from proximal, mid and distal regions and grown in the presence of ENR (d, e) or WENR (d, f). Scale bars, 100 μm. Results in c and d are representative of n = 3 independent experiments. In b, e and f data presented are means ± s.d. from n = 3 biologically independent samples. In e and f statistical significance was assessed using two-tailed Welch’s t-test.