Supplemental Figure 1

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1 Supplemental Figure 1 A B 50 * NeurofilamentM/βactin Relative expression control D neurogenic cond. PPARγ/ Relative expression C PPARγ Hprt OilRedO adipogenic cond. control 125 *

2 Supplemental Figure 2 rat dental epithelium (SF2 cells) phase (SF2) GFP pef6/gfppdgftmmycha selection SF2/GFP cocultured with cells SF2/GFP cells expressing GFPmycHA tag

3 Supplemental Figure 3 A B SF224 ips Clone No. Ambn Rat dental epithelium (SF2 cell) Ambn CK14 Klf4 Sox2 Oct3/4 Nanog

4 Supplemental Figure 4 A B m totalrna r totalrna (ng) (ng) mambn ips MEF tooth germ mambn rambn

5 Supplemental Figure 5 SF224iPS (days) Ambn Epiprofin/Sp6

6 SUPPLEMENTAL INFORMATION Supplemental Figure 1. Adipogenic and neurogenic induction of cells. (A) Adipocytespecific OilRedO staining of,, and cells in regular or adipogenic induction culture (adipogenic cond.) medium. (B) RTPCR and realtime PCR analyses of the adipocyte marker gene PPARγ in,, and cells in adipogenic induction culture medium. (C) Phase images of, and cells cultured in regular or neurogenic induction culture (neurogenic cond.) medium. (D) Western blotting of,, and cells cultured in regular () or neurogenic induction culture () medium was performed using the antineurofilamentm antibody. The relative intensities of bands obtained with western blots were scanned and quantified using an LAS UV4000 system (FujifilmGE Healthcare, Tokyo, Japan). Supplemental Figure 2. Schematic diagram of establishment of SF2 cells expressing PDGFtmGFPmycHA. Ratderived dental epithelial SF2 cells were transfected with a GFPPDGFtmmycHA expression vector and selected with antibiotics. GFP in the SF2 cells was visualized using fluorescence microscopy. SF2/GFP cells were cocultured with cells (bottom) and separated with the antiha antibody. Supplemental Figure 3. Gene expression of subcloned SF2 cell lines and ips cells. (A) Ambn expression in dental epithelial cells subcloned from SF2 cells. Clone SF224 expressed Ambn at the highest level and was used for coculturing with ips cells. (B) SF224 cells expressed the ameloblast marker genes CK14 and Ambn, but not stem cell markers. In contrast, stem cell markers were observed in ips cells, but not ameloblast markers. Supplemental Figure 4. Design of mousespecific Ambn primer. Mouse and rat Ambn mrna showed a high degree of homology (94.2%), thus regular Ambn primers for RTPCR are able to precisely distinguish between them. To resolve this problem, we employed a mouse Ambn LNA primer. The accuracy of the mousespecific Ambn LNA primer was confirmed by PCR with gradient cdna mixtures containing serial dilution ratios of mouse and/or rat cdnas. (A) The expected size of the amplicon using the mouse LNA primer was observed only with mouse cdna, while the intensity of the band increased in a cdna dosedependent manner. (B) Undifferentiated ips cells and MEF feeder cells did not express Ambn mrna. A mouse tooth germ sample served as a positive control for the mouse LNA primer. Supplemental Figure 5. Expression of epiprofin/sp6 transcription factor in ips cells cocultured with SF224 cells. Time course analyses of Ambn and epiprofin/sp6 gene expressions in ips cells cocultured with SF224 cells for 1 (1D), 7 (7D) and 10 days (10D) were analyzed by RTPCR.

7 Supplemental Table1 Gene name forward primer reverse primer Size bp (mouse/rat) mouse/rat Ambn 5 GCGTTTCCAAGAGCCCTGATAAC3 5 AAGAAGCAGTGTCACATTTCCTGG3 366/368 mouse/rat CK14 5 GAGAAGAACCGCAAGGATGC3 5 AGGTTATTCTCCAGGGATGCTTTC3 191 mouse/rat 5 GGAGCGAGACCCCACTAACATC3 5 CTCGTGGTTCACACCCATCAC3 181 mouse Ambn 5 GCAGGTGGCACCATCCGALA3 5 AAGAAGCAGTGTCACATTTCCTGG rat Ambn 5 GCTACCCTTCCACAGGGAAGGTLA3 5 AAGAAGCGGTGTCACATTTCCTGG3 267 mouse Enam 5 GGGGACCACCAACAGCGTTTGGACGGCCA3 5 CCCGGAGGTAGGAGAGGGAGGTTTACC3 614 rat Enam 5 CAGGACCACCAACAGGGTTCGGACGA3 5 TCTTGAGGTAGGAGGGGAAGGTTTGT3 626 mouse CK14 5 GGCTGCCGATGACTTCCGGAC3 5 CAGCAGTATCTGCGTCCACGCA3 909 rat CK14 5 AGACTACAGCCCCTACTTCAAGACC3 5 AGGTTATTCTCCAGGGATGCTTTC3 597 mouse p63 5 GGCATTTCAGCACTATTTAGGTGG3 5 CACTGGTGTGAGGAGACAAACTGTG3 110 mouse Epiprofin 5 TCAGCCTGCTTTGGGAGGATAC3 5 ATGCTTCTTCTTGCCCCCATCG3 313 Klf4 5 CACCATGGACCCGGGCGTGGCTGCCAGAAA3 5 TTAGGCTGTTCTTTTCCGGGGCCACGA3 739 Sox2 5 GGTTACCTCTTCCTCCCACTCCAG3 5 TCACATGTGCGACAGGGGCAG3 193 Oct3/4 5 TCTTTCCACCAGGCCCCCGGCTC3 5 TGCGGGCGGACATGGGGAGATCC3 224 Nanog 5 CAGGTGTTTGAGGGTAGCTC3 5 CGGTTCATCATGGTACAGTC3 223 Fgf4 5 CGTGGTGAGCATCTTCGGAGTGG3 5 CCTTCTTGGTCCGCCCGTTCTTA3 197 Gdf3 5 GTTCCAACCTGTGCCTCGCGTCTT3 5 AGCGAGGCATGGAGAGAGCGGAGCAG3 570 Cdx2 5 GGCGAAACCTGTGCGAGTGGATGCGGAA3 5 GATTGCTGTGCCGCCGCCGCTTCAGACC3 493/490 Gata6 5 ACCTTATGGCGTAGAAATGCTGAGGGTG3 5 CTGAATACTTGAGGTCACTGTTCTCGGG3 334/328 Brachyury 5 ATGCCAAAGAAAGAAACGAC3 5 AGAGGCTGTAGAACATGATT3 835 B2 5 GGGACCCGCTGTCTTCTAGT3 5 TCAACTCAAATTCGCTGAGGAC3 154 B4 5 ACTGCCGCAGCTTCTCTGAG3 5 TTCTCCAGATGTTCTTCGTG3 486 Osteocalcin 5 CCTCTTGAAAGAGTGGGCTG3 5 CCTCGGGAGACAAACAACAT3 268 Osteonectin 5 GTCTCACTGGCTGTGTTGGA3 5 AAGACTTGCCATGTGGGTTC3 264 Runx2 5 GACGTTCCCAAGCATTTCAT3 5 ACTCTGGCTTTGGGAAGAG3 181 Dspp 5'GGAACTGCAGCACAGAATGA3' 5'CAGTGTTCCCCTGTTCGTTT3' 199/193 Bcrp1 5' GGTGAATCTCAGAACCATTGGGC3 5'GCTGTTGTCCGTTACATTGAATCC 3' 139 PPARγ 5 CTGATGCACTGCCTATGAGC3 5 CAGACTCGGCACTCAATGGC3 373 Hprt 5 GTTAAGCAGTACAGCCCCAAA3 5 AGGGCATATCCAACAACAAACTT3 131 Gapdh (Realtime) 5 CCATCACCATCTTCCAGGAG3 5 GCATGGACTGTGGTCATGAG3 322