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. 2012 Dec 12;110(2):E141–E150. doi: 10.1073/pnas.1214361110

Fig. 2.

Fig. 2.

Inhibition of Hh signaling through cyclopamine (cyc) blocks hematopoietic development in EBs. (A) CFUs measured from day 6 whole EBs in response to cyc treatment during days 2–5 of differentiation (n = 4). (B) Flow cytometric quantification of CD41+c-Kit+ hematopoietic precursors cells from day 6 whole EBs in response to cyc treatment during days 2–5 of differentiation. Horizontal bars indicate mean values (n = 5). (C) Hematopoietic gene expression measured in day 6 whole EBs after cyc treatment from days 2 to 5 of differentiation as assessed by quantitative PCR (n = 3). (D) Gene expression time course of Hedgehog target Gli1 and mesoderm-related genes Brachyury and Cerberus during days 2–5 of cyc treatment using whole EBs (n = 2). P values were derived from one-way ANOVA for correlated samples. (E) Flk1+ levels (shown in red) on days 3.25–3.75 after cyc treatment from day 2 of differentiation as assessed by flow cytometry. Black lines represent the isotype controls (n = 2). (F) Percentage of beating EBs on days 7–8 in response to cyc treatment during days 2–5 of differentiation (n = 3). (G) Reduction in cardiac troponin T (cTnT) gene expression in whole EBs in response to cyc treatment from day 2 to 5 (n = 3). (H) Blast colony formation during treatment of day 3.5 Flk1+ with cyc for 4 d in BL-CFC media (n = 3). (I) Core colony formation during treatment of day 3.5 Flk1+ with cyc for 4 d in BL-CFC media (n = 3). (J) Diagram of cell subpopulations affected by cyclopamine treatment.