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sc-12-0798_sm_SupplFigure1.tif1574KSupplemental Fig. S1. Lgr5 expression in circumvallate papilla. Confocal images showing that the strongest-staining Lgr5-GFP cells (green) are adjacent to the opening of the duct of Von Ebner's gland below the circumvallate papilla. A weaker Lgr5-GFP signal is found in the basal cells or perigemmal cells at the base or just outside of the taste buds. Type II taste receptor cells were identified by anti-Trpm5 immunoreactivity (red). DAPI (blue) staining was used to visualize nuclei. d, duct of Von Ebner's gland. The square in (A) corresponds to the area shown at higher magnification in (B). Scale bars: a=80 μm; b=40 μm.
sc-12-0798_sm_SupplFigure2.tif1365KSupplemental Fig. S2. Lgr5 expression in foliate papillae. (A) Transmitted light image of foliate papillae. (B) Corresponding confocal image showing Lgr5-GFP cells (green) most strongly expressed in the trench area below the foliate papillae. A weaker Lgr5-GFP signal is found in the basal cells or perigemmal cells at the base or just outside of the taste buds. Type II taste receptor cells were identified by anti-Trpm5 immunoreactivity (red). d, duct of Von Ebner's gland. Scale bars: 40 μm.
sc-12-0798_sm_SupplFigure3.tif1541KSupplemental Fig. S3. Lgr5-expressing cells continue to generate mature taste bud cells 6 months after induction. (A and B) β-Galactosidase+ cells from the Lgr5+ lineage are found in circumvallate papilla taste bud cells as well as in perigemmal cells (white arrows) 6 months after 5-days of tamoxifen induction of Lgr5-Cre. In addition, β-galactosidase+ cells are found in the trench area at the opening of the Von Ebner's gland (black arrow). (C) 6 months after 5 days of tamoxifen induction many β-galactosidase-stained mucosal cells of the Lgr5+ lineage are present in crypts and villi of the small intestine. d, duct of Von Ebner's gland. Scale bars: 40 μm.
sc-12-0798_sm_SupplFigure4.tif2341KSupplemental Fig. S4. Lgr5-expressing cells generate mature taste cells after a single tamoxifen injection. (A, B) One day after 1-day single tamoxifen injection, a small number of β-galactosidasestained taste cells (blue) of the Lgr5+ lineage are present only at the bottom of the trench area (black arrows). β-Galactosidase+ cells from the Lgr5+ lineage are found in circumvallate papilla taste bud cells (white arrows) as well as in perigemmal cells (arrowheads) 2 weeks (C, D) and 1 month (E, F) after a single tamoxifen injection. Scale bars: 20 μm.
sc-12-0798_sm_SupplFigure5.tif1526KSupplemental Fig. S5. Lgr5-expressing cells generate mature taste cells in the foliate papillae 1 month after a single tamoxifen injection. Tamoxifen-induced Lgr5-Cre generates β-galactosidase activity to mark cells from the Lgr5+ lineage. Intragemmal and perigemmal β-galactosidase+ cells (blue) are found in foliate papillae taste buds (arrows). A and B are two representative sections. Scale bars: 20 μm.
sc-12-0798_sm_SupplFigure6.pdf795KSupplemental Fig. S6. Lgr5+ stem cells generate all three types of taste cells. Mature taste bud cells marked by Lgr5-Cre-generated β-galactosidase 1-2 months after tamoxifen induction were double stained with anti-β-galactosidase and markers for each of the three types of taste cells. A-D) β-Galactosidase immunoreactivity (green) is co- expressed in type I taste cells with NTPDase2 (red), two months after a single tamoxifen induction. E-H) Lgr5-Cre-generated β-galactosidase immunoreactivity (green) is coexpressed in type II taste receptor cells with Trpm5 (red), 2 months after 5-days of tamoxifen injections. I-L) Lgr5-Cre-generated β-galactosidase immunoreactivity (green) is co-expressed in type III taste receptor cells with serotonin (red), six weeks after a single tamoxifen injection. All sections are compressed Z-stacks (4-10 μm thickness). Scale bars: A, E, and I=20 μm; B-D, F-H, and J-L=8 μm.
sc-12-0798_sm_SupplFigure7.tif1546KSupplemental Fig. S7. Mature taste cells generated from Lgr5-expressing cells 6 months after induction. Lgr5-Cre-generated β-galactosidase immunoreactivity (green) generated 6 months after induction by 5-days of tamoxifen is co-expressed with Trpm5 (red) in type II taste receptor cells. Asterisks indicate colocalization of β-galactosidase and Trpm5. (A and B) show two representative dual-stained sections. Scale bars: 20 μm.
sc-12-0798_sm_SupplTable1.pdf28KSupplementary Table 1

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