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. 2020 Oct 27;147(20):dev195388. doi: 10.1242/dev.195388

Fig. 1.

Fig. 1.

Ror2 is expressed in both midgut epithelium and mesenchyme, but epithelial Ror2 is dispensable for gut elongation. (A-E) Immunostaining of ROR2 (white) on cross-sections of the central wild-type midgut at E10.5-E14.5. Scale bars: 20 µm. (F-H′) Immunostaining of ROR2 (white) and fluorescence of membrane tdTomato (magenta) on E14.5 midgut sections. (G-H′) Higher magnifications of boxed epithelial (G,G′) and mesenchymal (H,H′) regions in F. In G′, the yellow dashed line outlines the apical surface. Scale bars: 20 µm. (I-K″) RNAscope in situ hybridization with probes for Ror2 on cross-sections of the central midgut at E10.5, E12.5 and E14.5. (K′,K″) Higher magnifications of the boxed epithelial (K′) and mesenchymal (K″) regions in K. The epithelial-mesenchymal interface is outlined by dashed white lines. Scale bars: 20 µm. (L-O) E14.5 embryos and isolated gastrointestinal (GI) tracts of the control (L,M) and epithelial Ror2 knockout (N,O). Scale bars: 1 mm. (P-S) Immunostaining of ROR2 (white) and fluorescence of membrane tdTomato (magenta) and membrane eGFP (mG, green) on cross-sections of control (P,Q) and epithelial Ror2 knockout (R,S) midgut at E14.5. Scale bars: 20 µm. (T,U) Quantitation of midgut length (from the pylorus to the cecum) and crown-rump length of embryos at E14.5 (T) and E18.5 (U) with (gray) or without (green) epithelial ROR2. At E14.5, Ror2flox/+; ShhCre/+, n=3; Ror2flox/flox and Ror2flox/+, n=6; Ror2flox/flox; ShhCre/+, n=4. At E18.5, Ror2flox/+; ShhCre/+, n=4; Ror2flox/flox and Ror2flox/+, n=12; Ror2flox/flox; ShhCre/+, n=4. (V-X) Quantitation of basal circumference, epithelial height and nuclei number on cross-sections of the central midgut epithelial tubes with or without epithelial ROR2. Quantification was performed on 25 sections of five control samples and 20 sections of four Ror2flox/flox; ShhCre/+ samples. For the basal circumference and nuclei number, one measurement was taken per section; for the epithelial height, three or four measurements were taken per section. Data are mean±s.e.m. Analyses were performed using unpaired nonparametric tests (Mann–Whitney test). n.s., not significant.