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Additional Supporting Information may be found in the online version of this article.

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sc-12-0816_sm_SupplFigure1.pdf1752KFigure S1. H9 cells were cultured on Matrigel and treated with 1 μM BIO for 3 days before exposure to 100 ng/ml Activin A at day 0 and 5 ng/ml BMP4 at day 1 in RPMI/B27 with or without insulin. 30 days post-initiation of differentiation, cells were analyzed for cTnT expression by flow cytometry.
sc-12-0816_sm_SupplFigure2.tif2560KFigure S2. H9 cells were cultured on Matrigel and treated with 1 μM BIO for 3 days before exposure to 100 ng/ml Activin A at day 0 and 5 ng/ml BMP4 at day 1 in RPMI/B27 with or without insulin. 7 days post-initiation of differentiation, cells were analyzed for brachyury and Nkx2.5 expression by flow cytometry. Error bars represent the s.e.m. of three independent experiments.
sc-12-0816_sm_SupplFigure3.tif2559KFigure S3. 19-9-11 cells were cultured on Matrigel in mTeSR1 and when the monolayer of cells reached 80- 90% confluence, a thin layer of Matrigel (0.5 mg per 6-well plate) was overlaid. Cells were then cultured in mTeSR1 for another 1-2 days until the cells were 100% confluent. At this point, day 0, the cells were exposed to 100 ng/ml Activin A and Matrigel (0.5 mg/6-well plate) in RPMI/B27 with or without insulin. After 24 hours, the medium was changed and cells were exposed to 10 ng/ml BMP4 and 10 ng/ml bFGF in RPMI/B27 with or without insulin. 15 days post-initiation of differentiation, cells were analyzed for cTnT expression by flow cytometry. Error bars represent the s.e.m for at least two independent experiments.
sc-12-0816_sm_SupplFigure4.tif2742KFigure S4. H9-7TGP-ishcat-1 cells were differentiated as described in Fig. 5(A). 10 μg/ml insulin and 2 μg/ml dox was added or not, as indicated, into the culture medium at day 2. 15 days after initiation of differentiation, cells were analyzed by flow cytometry using the MF20 antibody. Error bars represent the s.e.m. of three independent experiments.
sc-12-0816_sm_SupplFigure5.tif2662KFigure S5. H9 cells were cultured on Matrigel and treated with 1 μM BIO for 3 days before exposure to 100 ng/ml Activin A at day 0 and 5 ng/ml BMP4 at day 1 in RPMI/B27 with or without insulin (GiAB protocol). H9 cells were also differentiated as described in Fig. 4(A) (GiWi protocol). 30 days post-initiation of differentiation, cells were analyzed for PECAM-1 and smooth muscle actin (SMA) expression by flow cytometry. Error bars represent the s.e.m. of three independent experiments.
sc-12-0816_sm_SupplMovie1.mpg2716KMovie S1. H9 cells were differentiated according to the GiAB protocol shown in Fig. 1(A). RPMI/B27 without insulin was used for differentiation. Movie S1 shows day 15 cardiomyocytes.
sc-12-0816_sm_SupplMovie2.mpg1696KMovie S2. H9 cells were differentiated according to the GiAB protocol shown in Fig. 1(A). RPMI/B27 containing insulin was used for differentiation. Movie S2 shows day 15 differentiated cells.
sc-12-0816_sm_SupplMovie3.mpg4356KMovie S3-5. H9 cells were differentiated as described in Movie S1 with RPMI/B27-insulin as basal medium. At day 1, 0 μg/ml (Movie S3), 1 μg/ml (Movie S4) or 10 μg/ml (Movie S5) insulin was added into the cell culture. Movies show day 15 differentiated cells.
sc-12-0816_sm_SupplMovie4.mpg2114KMovie S3-5. H9 cells were differentiated as described in Movie S1 with RPMI/B27-insulin as basal medium. At day 1, 0 μg/ml (Movie S3), 1 μg/ml (Movie S4) or 10 μg/ml (Movie S5) insulin was added into the cell culture. Movies show day 15 differentiated cells.
sc-12-0816_sm_SupplMovie5.mpg2498KMovie S3-5. H9 cells were differentiated as described in Movie S1 with RPMI/B27-insulin as basal medium. At day 1, 0 μg/ml (Movie S3), 1 μg/ml (Movie S4) or 10 μg/ml (Movie S5) insulin was added into the cell culture. Movies show day 15 differentiated cells.
sc-12-0816_sm_SupplTable1.tif2121KSupplementary Table 1
sc-12-0816_sm_SupplTable2.tif2760KSupplementary Table 2

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