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. 2014 Apr 24;3:e02164. doi: 10.7554/eLife.02164

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Copyright © 2014, Shenje et al

This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.

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Figure 6. — (A–G′) Confocal images of PA2 sections of control (A–C‴), Numb/Numbl DKO (D), and chimeric (E–G′) embryos, immunostained with PH3, RFP, Isl1 antibodies. Asterisks indicate PH3⁺ RFP⁺ Isl1⁺ (triple positive) cells located in outer layers of RFP⁺ Isl1⁺ cell population (outlined, A and E). Boxed areas in (A), (B), and (E) are shown in higher magnification in (B), (C–C‴), and (F–G′), respectively. No PH3⁺ cells are found in RFP⁺ cells (asterisks, F–G′). RFP⁺ cells were outlined in white (F–G′). (H and I) Percentage of donor-derived Isl1⁺ Nkx2.5⁻ cells in PA2 is shown in (H) and number of PH3⁺ RFP⁺ Isl1⁺ cells per 12-micron PA2 section is shown (I). Data are mean ± SD; n = 10; *p<0.05. (J and K) Relative percentage of EdU⁺ cells in ES cell-derived Mesp1⁺ progenitor, Isl1⁺ Nkx2.5⁻ CPCs or Nkx2.5⁺ CPCs transfected with control or Numb/Numbl DKO siRNA (J) or Isl1⁺ Nkx2.5⁻ CPCs transfected with control (CTV) or Numb overexpression construct (CTV-Numb) (K). Data are mean ± SD; n = 3; *p<0.05. The Mesp1⁺, Isl1⁺ Nkx2.5⁻, or Nkx2.5⁺ cells were FACS-purified from day 4 Mesp1^Cre; Ai9, day 5 Isl1^Cre; Ai9, or day 6 Nkx2.5^GFP ES cells, respectively. Dapi (blue) was used to counterstain the nuclei. p values were determined using the paired Student t test. Scale bars, 10 μm (B, C, F, G). 50 μm (A, D, E). fe, foregut endoderm; pa, pharyngeal arch ec, endocardial layer; mc, myocardial layer; pc, pericardial layer.

DOI: http://dx.doi.org/10.7554/eLife.02164.021

Figure 6—figure supplement 1. — (A–G′) Confocal images of PA2 sections of control (A–C‴), Numb/Numbl DKO (D), and chimeric (E–G′) embryos, immunostained with PH3, RFP, Isl1 antibodies. Asterisks indicate PH3⁺ RFP⁺ Isl1⁺ (triple positive) cells located in outer layers of RFP⁺ Isl1⁺ cell population (outlined, A and E). Boxed areas in (A), (B), and (E) are shown in higher magnification in (B), (C–C‴), and (F–G′), respectively. No PH3⁺ cells are found in RFP⁺ cells (asterisks, F–G′). RFP⁺ cells were outlined in white (F–G′). (H and I) Percentage of donor-derived Isl1⁺ Nkx2.5⁻ cells in PA2 is shown in (H) and number of PH3⁺ RFP⁺ Isl1⁺ cells per 12-micron PA2 section is shown (I). Data are mean ± SD; n = 10; *p<0.05. (J and K) Relative percentage of EdU⁺ cells in ES cell-derived Mesp1⁺ progenitor, Isl1⁺ Nkx2.5⁻ CPCs or Nkx2.5⁺ CPCs transfected with control or Numb/Numbl DKO siRNA (J) or Isl1⁺ Nkx2.5⁻ CPCs transfected with control (CTV) or Numb overexpression construct (CTV-Numb) (K). Data are mean ± SD; n = 3; *p<0.05. The Mesp1⁺, Isl1⁺ Nkx2.5⁻, or Nkx2.5⁺ cells were FACS-purified from day 4 Mesp1^Cre; Ai9, day 5 Isl1^Cre; Ai9, or day 6 Nkx2.5^GFP ES cells, respectively. Dapi (blue) was used to counterstain the nuclei. p values were determined using the paired Student t test. Scale bars, 10 μm (B, C, F, G). 50 μm (A, D, E). fe, foregut endoderm; pa, pharyngeal arch ec, endocardial layer; mc, myocardial layer; pc, pericardial layer.

DOI: http://dx.doi.org/10.7554/eLife.02164.021