Figure S1 The effect of FGF-10 in controlling ES cell differentiation. A, The percentage of cardiomyocyte differentiation of ES cell treated with FGF-10 over time from day 7 to day 14. B, the time course study of the three germ layer markers upon FGF- 10 treatment with quantitative real-time PCR. Found at: doi:10.1371/journal.pone.0014414.s002 (6.68 MB TIF) Figure S2 The study of ES cell toxicity under the treatment of FGF receptor inhibitor PD173074. A, Immunoblot analysis of phospho- and total-Akt in embryoid bodies of different treatment. FGF-10, 100 ng/ml. Neutralizing antibody, 0.1mg/ml. PD173074, 30 nM. Control, no treatment. B-D, Total cell count of ES cells treated with an FGF-10 neutralizing antibody, the FGFR inhibitor PD173074, or a lentivirus contained FGF -10, FGFR-1 or FGFR- 2 shRNA. Control, no treatment. Found at: doi:10.1371/journal.pone.0014414.s003 (4.26 MB TIF) Figure S3 Gene expression profiles of ES cells treated with FGF-10 protein or shRNA. A, RT-PCR results showing the temporal expression pattern of Brachyury, Flk1, Gata4, aMHC, cTnI and cTnT on day 3 (D3), 5 (D5), 7 (D7) and 9 (D9) during hanging drop cultivation of ES cells after the treatment of 100 ng/ml FGF-10. (n§3, *p , 0.05, **p , 0.01, ***p , 0.001.). B, RT-PCR results showing the temporal expression pattern of Brachyury, Flk1, Gata4, aMHC, cTnI and cTnT on day 3 (D3), 5 (D5), 7 (D7) and 9 (D9) during hanging drop cultivation of ES cells after FGF-10 knockdown by shRNA (n§3, *p , 0.05, **p , 0.01, ***p , 0.001.). References 1. Jessup M, Brozena S (2003) Heart failure. N Engl J Med 348: 2007–2018. 2. Writing Group Members, Rosamond W, Flegal K, Furie K, Go A, Greenlund K, et al. (2008) Heart disease and stroke statistics—2008 update: a report from the American Heart Association Statistics Committee and Stroke Statistics Subcommittee. Circulation 117: e25–146. 3. Quevedo HC, Hatzistergos KE, Oskouei BN, Feigenbaum GS, Rodriguez JE, et al. 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FGF10 Cardiac Differentiation Found at: doi:10.1371/journal.pone.0014414.s004 (3.53 MB TIF) Video S1 Bright field image of control with no treatment Found at: doi:10.1371/journal.pone.0014414.s005 (1.06 MB AVI) Video S2 Fluorescence image of control with no treatment Found at: doi:10.1371/journal.pone.0014414.s006 (0.91 MB AVI) Video S3 Bright field image of embryoid bodies treated with 100 ng/ml of FGF-10 Found at: doi:10.1371/journal.pone.0014414.s007 (0.96 MB AVI) Video S4 Fluorescence image of embryoid bodies treated with 100 ng/ml of FGF-10 Found at: doi:10.1371/journal.pone.0014414.s008 (1.39 MB AVI) Acknowledgments We greatly appreciate Iain Bruce (Zhejiang University) for manuscript preparation and Judy Wang, Katie Wu and Ting-Yu Chu (National Cheng Kung University) for technical support. Author Contributions Conceived and designed the experiments: SSKC HJL YCH JHC SMH PCHH. Performed the experiments: SSKC HJL YCH DSL CYC ES. Analyzed the data: SSKC HJL YCH DSL PCHH. 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Hsieh PCH, Davis ME, Gannon J, MacGillivray C, Lee RT (2006) Controlled delivery of PDGF-BB for myocardial protection using injectable self-assembling peptide nanofibers. J Clin Invest 116: 237–248. PLoS ONE | www.plosone.org 12 December 2010 | Volume 5 | Issue 12 | e14414