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. Author manuscript; available in PMC: 2015 May 28.
Published in final edited form as: Nat Commun. 2013;4:1367. doi: 10.1038/ncomms2378

Figure 3.

Figure 3

Differentiation of hiPSCs into the intermediate mesoderm (IM). (a) Flow cytometric analyses of OSR1(GFP)+ cell differentiation on culture day 11. 1: no factors; 2: 100 ng/ml BMP7; 3: Stage 1, 100 ng/ml activin A + 100 ng/ml Wnt3a, Stage 2, 100 ng/ml BMP7. (b) The differentiation of OSR1(GFP)+ cells on culture day 11, analyzed by flow cytometry. 1: on day 1 before treatments; 2: no factors; 3-7: Stage 1, 100 ng/ml activin A + 100 ng/ml Wnt3a, The treatments at stage 2 are as follows, 3: 100 ng/ml Wnt3a; 4: 100 ng/ml BMP7; 5: 100 ng/ml BMP7 + 25 ng/ml Wnt3a; 6: 10 ng/ml BMP7 + 100 ng/ml Wnt3a; 7: 100 ng/ml BMP7 + 100 ng/ml Wnt3a. (c) The suppression of the induction efficiency of OSR1(GFP)+ cells on culture day 11 by adding Frizzled- and/or Noggin-Fc chimeric proteins to the treatment with 100 ng/ml BMP7 + 100 ng/ml Wnt3a at Stage 2. The treatment at Stage 1 is 100 ng/ml activin A + 100 ng/ml Wnt3a in all experiments. 1: without Frizzled- or Noggin-Fc; 2: 1 mg/ml Frizzled-Fc; 3: 1 mg/ml Noggin-Fc; 4: 1 mg/ml Frizzled-Fc + 1 mg/ml Noggin-Fc. The data from three independent experiments are presented as the means ± SD (n=3).