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. 2013 Feb 4;110(8):3023–3028. doi: 10.1073/pnas.1217039110

Fig. 4.

Fig. 4.

Effects of Wnt3a in combination with high-dose G-CSF. (A) Filled and open bars show the proportion of mature neutrophils among the cells derived from SCN-iPS cells (SPN0101) on day 14 of suspension culture in the presence of neutrophil differentiation medium containing 10 ng/mL G-CSF (GCSF 10 ng/mL); 500 ng/mL Wnt3a and 10 ng/mL G-CSF (Wnt3a+GCSF 10 ng/mL); 1,000 ng/mL G-CSF (GCSF 1,000 ng/mL); or 500 ng/mL Wnt3a and 1,000 ng/mL G-CSF (Wnt3a + GCSF 1,000 ng/mL); and that from control iPS cells (controls 1 and 2) cultured in the neutrophil differentiation medium containing 10 ng/mL G-CSF, respectively. Data are shown as mean ± SD. ★★P < 0.01; ★P < 0.05. (B) The proportion of mature neutrophils among the cells derived from three SCN-iPS cell clones (SPN0101, SPN0102, and SPN0103) on day 14 of suspension culture in the presence of neutrophil differentiation medium containing 10 ng/mL G-CSF (GCSF 10 ng/mL); 500 ng/mL Wnt3a and 10 ng/mL G-CSF (Wnt3a+GCSF 10 ng/mL); or 500 ng/mL Wnt3a and 1,000 ng/mL G-CSF (Wnt3a + GCSF 1,000 ng/mL). Data are shown as mean ± SD. ★★P < 0.01; ★P < 0.05. (C) Filled and open bars show the relative expression (target/HPRT expression) of LEF-1 mRNA in SCN-iPS cells (SPN0101) on day 2 of suspension culture in the presence of differentiation medium containing the same combinations of Wnt3a and G-CSF as shown in A and that from control iPS cells (control 2), respectively. Data are shown as mean ± SD. ★★P < 0.01; ★P < 0.05. (D) Filled and open bars show the relative expression (target/HPRT expression) of C/EBP-α, BIRC5, or cyclin D1 mRNA in SCN-iPS cells (SPN0101) on day 2 of suspension culture in the presence of differentiation medium containing the same combinations of Wnt3a and G-CSF as shown in A and that from control iPS cells (control 2), respectively. Data are shown as mean ± SD. ★★P < 0.01; ★P < 0.05. (E) Total cell numbers of nonadherent cells in suspension cultures of 1 × 104 CD34+ cells derived from control iPS cells (control 2; red broken line) and SCN-iPS cells (SPN0101) in the presence of neutrophil differentiation medium (black line) and those from SCN-iPS cells in the presence of neutrophil differentiation medium containing 500 ng/mL Wnt3a (yellow line) or 1,000 ng/mL G-CSF (black line). Data are shown as mean ± SD. ★★P < 0.05.