Figure 4
Figure 4. Suppression of erythropoiesis by miRs-126/126*. (A) A total of 1 μg/mL Dox treatment induced expression of miRs-126/126* in differentiating EBs generated from I-miRs-126/126* hESCs. miR expression is presented as fold induction relative to expression in day 10 EB cells without Dox treatment. (B) Phase-contrast and fluorescence microscopic images taken from representative erythroid (E), macrophage (M), granulocyte-macrophage (GM), and mixed myelo-erythroid (GEMM) colonies derived from day 15 miRs-126/126* overexpressing EB cells. Images of E and GEMM colonies are 100× and of M and GM colonies, 50×. Wright-Geimsa images are 200×. Phase contrast and fluorescent micrographs of live cells in methylcellulose were acquired using a Zeiss Axiovert 200 model microscope with ECPlan Neofluor 5×/0.16 or 10×/0.3 Ph1 objectives. Wright-Geimsa–stained cytospins were imaged using a Nikon Eclipse E600 model microscope with PlanApo 20×/0.75 Ph2 DM objectives. Image was captured using an Optronics MagnaFIRE camera with MicroFIRE Version 070121-00X1 software. (C) Total number of hematopoietic CFU formed in methylcellulose. (D) CFU-E output of day 15 EB cells generated from different Dox treatments. (E) Appearance of the pellets of colonies grown in methylcellulose medium for 12 days. Data are mean ± SD (n = 4). *P < .05. **P < .01

Suppression of erythropoiesis by miRs-126/126*. (A) A total of 1 μg/mL Dox treatment induced expression of miRs-126/126* in differentiating EBs generated from I-miRs-126/126* hESCs. miR expression is presented as fold induction relative to expression in day 10 EB cells without Dox treatment. (B) Phase-contrast and fluorescence microscopic images taken from representative erythroid (E), macrophage (M), granulocyte-macrophage (GM), and mixed myelo-erythroid (GEMM) colonies derived from day 15 miRs-126/126* overexpressing EB cells. Images of E and GEMM colonies are 100× and of M and GM colonies, 50×. Wright-Geimsa images are 200×. Phase contrast and fluorescent micrographs of live cells in methylcellulose were acquired using a Zeiss Axiovert 200 model microscope with ECPlan Neofluor 5×/0.16 or 10×/0.3 Ph1 objectives. Wright-Geimsa–stained cytospins were imaged using a Nikon Eclipse E600 model microscope with PlanApo 20×/0.75 Ph2 DM objectives. Image was captured using an Optronics MagnaFIRE camera with MicroFIRE Version 070121-00X1 software. (C) Total number of hematopoietic CFU formed in methylcellulose. (D) CFU-E output of day 15 EB cells generated from different Dox treatments. (E) Appearance of the pellets of colonies grown in methylcellulose medium for 12 days. Data are mean ± SD (n = 4). *P < .05. **P < .01

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