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Files in this Data Supplement:
Fig. S1. Fgf9, desmin and p-Smad1/5/8 expression in embryonic mouse small intestines. (A) Whole-mount small intestine from a wild-type E14.5 mouse used for in situ hybridization using a probe for Fgf9. The staining is consistent with epithelial expression, as was demonstrated at earlier time points (Zhang et al., 2006). (B-E) Immunofluorescence-stained sections of E18.5 small intestines from (B,D) control and (C,E) Fgf9−/− embryos. Sections were stained with antisera directed against desmin (B,C; green) and p-Smad1/5/8 (D,E; red). There were no differences in desmin or p-Smad 1/5/8 expression when comparing small intestines from control and Fgf9−/− animals. Scale bars: 30 µm.
Fig. S2. p-Erk positive mesenchymal cells are diminished in DCR1R2 embryos and express Fst and Fgfr2. (A) Quantification of the mean percentage (±s.d.) of mesenchymal cells with detectable p-Erk in control and DCR1R2 embryos (E18.5). The double asterisk indicates a statistically significant difference when comparing DCR1R2 embryos with controls as determined by a Student’s t-test (P<0.01). Sections from control (B) E14.5 and (C,D) E18.5 small intestines stained with anti-Fgfr2 and (C) anti-pErk1/2 (red) and (D) anti-Fst (red). The broken white lines in B-D indicate the epithelial/mesenchymal junction. The broken yellow line in B indicates the mesenchymal/muscularis propria border. The cells circled by the broken yellow lines in C and D show mesenchymal cells that are positive for both Fgfr2 and p-Erk1/2 (C) and Fst (D). Scale bars: 5 µm.
Fig. S3. iMSCs are multipotent, Fgf9 responsive, and co-express Fst and pErk1/2. (A) Osteogenic induction of cultured iMSCs stained with alizarin red. The widespread positive cells indicated that iMSCs can differentiate into osteocytes. (B) Adipogenic induction of iMSCs stained with Oil Red O. The widespread positive cells indicated that iMSCs can differentiate into adipocytes. (C) Cultured, serum starved iMSCs phosphorylated Erk1/2 in response to treatment with 10 ng/ml Fgf9. Immunoblots at various time points post treatment are shown. (D) Cultured iMSC stained with antisera directed against p-Erk1/2 (green) and Fst (red). Arrow indicates the cell nucleus. Scale bars: 50 µm in A,B; 5 µm in D.
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