Figure 1.
Figure 1. p110α-deficient embryos develop Tie2-/-–like vascular defects at E9.5. (A-B) Yolk sacs. Arrowhead points to a major branch of vitelline vessels. (C-D) Embryos proper. Arrowheads indicate blood congestion. (E-F) Mid-trunk region of Pik3ca+/- and Pik3ca-/- embryos. White arrowheads point to ACV (E) or to the corresponding position in the mutant at which the ACV is poorly assembled (F). Red arrowheads point to dorsal aortae. Images are focused at the level of the ACV. (G-H) Transverse sections at the level of branchial arches. Arrowheads point to ACV lumens. (I-J) X-gal–stained hearts. (K-L) Transverse sections through the center of the heart. Black arrowheads point to endocardial linings. (M-N) Whole-mount, X-gal–stained yolk sacs. Arrowhead in panel M indicates a large vessel, and arrow points to a small branch. (O-P) Histologic sections of X-gal–stained yolk sacs. Arrowhead in panel O points to a large vascular lumen. (Q-R) Transverse sections at the forebrain region of X-gal–stained embryos. Note that vascular lumens in panel R are significantly dilated and that endothelial cells are only weakly stained (arrowhead). (S-T) X-gal–stained embryos at E9.5. Arrowhead in panel T indicates weak X-gal staining signal. Images for whole-mount tissues (A-F, I, J, M, N, S, and T) were taken with a CoolSNAP CCD digital camera (Photometrics-Roger Scientific, Tucson, AZ) attached to a Leica MZFLIII stereomicroscope (Leica Microsystems, Heerbrugg, Switzerland). A single objective lens (Leica Plan 1.0, 10× magnification, Leica Microsystems) was used to obtain the images. Openlab 3.0 (Improvision, Lexington, MA) was used for initial image acquisition, and Photoshop 5.0 (Adobe Systems, San Jose, CA) was used for subsequent processing. Images for histologic sections (G, H, K, L, O, and P-R) were photographed using Carl Zeiss manufactured images products (Carl Zeiss, Göttingen, Germany), including an AxioCam CR color digital camera, AxioCam image acquisition software, a Zeiss Apochromat objective lens (63× for G, H, K, L, Q, and R; 20× for O and P), and an AxioPlan2 binocular microscope. Images in panels A-D were taken from freshly dissected embryos without staining; other specimens were stained with X-gal. All whole-mount embryo/yolk sac photographs were taken with specimens completely submerged in phosphate-buffered saline. Histological sections were counterstained with nuclear fast red. Scale bars: (A-D) 1 mm; (E-F, M-N) 400 μm; (G-H, K-L) 60 μm; (I-J) 250 μm; (O-P) 120 μm; (Q-R) 50 μm; (S-T) 500 μm.

p110α-deficient embryos develop Tie2-/-–like vascular defects at E9.5. (A-B) Yolk sacs. Arrowhead points to a major branch of vitelline vessels. (C-D) Embryos proper. Arrowheads indicate blood congestion. (E-F) Mid-trunk region of Pik3ca+/- and Pik3ca-/- embryos. White arrowheads point to ACV (E) or to the corresponding position in the mutant at which the ACV is poorly assembled (F). Red arrowheads point to dorsal aortae. Images are focused at the level of the ACV. (G-H) Transverse sections at the level of branchial arches. Arrowheads point to ACV lumens. (I-J) X-gal–stained hearts. (K-L) Transverse sections through the center of the heart. Black arrowheads point to endocardial linings. (M-N) Whole-mount, X-gal–stained yolk sacs. Arrowhead in panel M indicates a large vessel, and arrow points to a small branch. (O-P) Histologic sections of X-gal–stained yolk sacs. Arrowhead in panel O points to a large vascular lumen. (Q-R) Transverse sections at the forebrain region of X-gal–stained embryos. Note that vascular lumens in panel R are significantly dilated and that endothelial cells are only weakly stained (arrowhead). (S-T) X-gal–stained embryos at E9.5. Arrowhead in panel T indicates weak X-gal staining signal. Images for whole-mount tissues (A-F, I, J, M, N, S, and T) were taken with a CoolSNAP CCD digital camera (Photometrics-Roger Scientific, Tucson, AZ) attached to a Leica MZFLIII stereomicroscope (Leica Microsystems, Heerbrugg, Switzerland). A single objective lens (Leica Plan 1.0, 10× magnification, Leica Microsystems) was used to obtain the images. Openlab 3.0 (Improvision, Lexington, MA) was used for initial image acquisition, and Photoshop 5.0 (Adobe Systems, San Jose, CA) was used for subsequent processing. Images for histologic sections (G, H, K, L, O, and P-R) were photographed using Carl Zeiss manufactured images products (Carl Zeiss, Göttingen, Germany), including an AxioCam CR color digital camera, AxioCam image acquisition software, a Zeiss Apochromat objective lens (63× for G, H, K, L, Q, and R; 20× for O and P), and an AxioPlan2 binocular microscope. Images in panels A-D were taken from freshly dissected embryos without staining; other specimens were stained with X-gal. All whole-mount embryo/yolk sac photographs were taken with specimens completely submerged in phosphate-buffered saline. Histological sections were counterstained with nuclear fast red. Scale bars: (A-D) 1 mm; (E-F, M-N) 400 μm; (G-H, K-L) 60 μm; (I-J) 250 μm; (O-P) 120 μm; (Q-R) 50 μm; (S-T) 500 μm.

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