Journal: eLife
Article Title: Vascular endothelial-specific loss of TGF-beta signaling as a model for choroidal neovascularization and central nervous system vascular inflammation
doi: 10.7554/eLife.107018
Figure Lengend Snippet: ( A ) COL4 immunostaining of control and Cdh5CreER;Tgfbr1 CKO/- flatmount retinas showing arteries, veins, and capillaries in the control retina and a high density of vascular tufts in the Cdh5CreER;Tgfbr1 CKO/- retina. ( B ) False color images from the indicated genotypes and ages showing a stacked Z-series of flatmount retinas color-coded by the depth of the vasculature. For control retinas, the blue-green-red color scheme corresponds to the inner two-thirds of the retina: blue, vitreal surface; green, ganglion cell layer and inner plexiform layer; and red, inner nuclear layer and outer plexiform layer. For mutant retinas, the blue-green-red color scheme corresponds to a shallower depth, as the most deeply penetrating vascular tufts go only as far as the inner edge of the inner nuclear layer: blue, vitreal surface; green, ganglion cell layer; and red, inner plexiform layer. Left column of three panels: postnatal day (P)14 control retina (upper image; white arrows point to tip cells in the inner plexiform layer [IPL]) and two regions from a P14 Cdh5CreER;Tgfbr1 CKO/- retina (lower). Center column of three panels: P26 control retina (upper) and two regions from a P26 Cdh5CreER;Tgfbr1 CKO/- retina (lower). Right column of three panels, ~P30 Ndp KO retina (upper) and two regions from an ~P30 Fzd4 -/- retina (lower). All images are at the same magnification and are from the midperiphery of the retina. ( C ) Flatmounts of P14 control, Ndp KO , and Cdh5CreER;Tgfbr1 CKO/- retinas showing CLDN5 and PLVAP immunostaining and Sulfo-NHS-biotin accumulation (detected with fluorescent streptavidin). ( D ) Quantification of Sulfo-NHS-biotin in the retinal parenchyma. Pixel intensities for territories between vascular segments were quantified from four flatmount retinas for each of the three genotypes (13 territories per retina). For the two cohorts of control and mutant retinas, all pixel intensity values were scaled so that the mean values from the control retinas equal 1.0. Scale bar in ( A ), 1 mm. Scale bar in ( B ), 100 µm. Scale bar in ( C ), 100 µm.
Article Snippet: Texas Red Streptavidin was from Vector Laboratories (SA-5006).
Techniques: Immunostaining, Control, Mutagenesis
Vector Laboratories is a verified supplier 
Vector Laboratories manufactures this product 
![( A ) COL4 immunostaining of control and Cdh5CreER;Tgfbr1 CKO/- flatmount retinas showing arteries, veins, and capillaries in the control retina and a high density of vascular tufts in the Cdh5CreER;Tgfbr1 CKO/- retina. ( B ) False color images from the indicated genotypes and ages showing a stacked Z-series of flatmount retinas color-coded by the depth of the vasculature. For control retinas, the blue-green-red color scheme corresponds to the inner two-thirds of the retina: blue, vitreal surface; green, ganglion cell layer and inner plexiform layer; and red, inner nuclear layer and outer plexiform layer. For mutant retinas, the blue-green-red color scheme corresponds to a shallower depth, as the most deeply penetrating vascular tufts go only as far as the inner edge of the inner nuclear layer: blue, vitreal surface; green, ganglion cell layer; and red, inner plexiform layer. Left column of three panels: postnatal day (P)14 control retina (upper image; white arrows point to tip cells in the inner plexiform layer [IPL]) and two regions from a P14 Cdh5CreER;Tgfbr1 CKO/- retina (lower). Center column of three panels: P26 control retina (upper) and two regions from a P26 Cdh5CreER;Tgfbr1 CKO/- retina (lower). Right column of three panels, ~P30 Ndp KO retina (upper) and two regions from an ~P30 Fzd4 -/- retina (lower). All images are at the same magnification and are from the midperiphery of the retina. ( C ) Flatmounts of P14 control, Ndp KO , and Cdh5CreER;Tgfbr1 CKO/- retinas showing CLDN5 and PLVAP immunostaining and Sulfo-NHS-biotin accumulation (detected with fluorescent <t>streptavidin).</t> ( D ) Quantification of Sulfo-NHS-biotin in the retinal parenchyma. Pixel intensities for territories between vascular segments were quantified from four flatmount retinas for each of the three genotypes (13 territories per retina). For the two cohorts of control and mutant retinas, all pixel intensity values were scaled so that the mean values from the control retinas equal 1.0. Scale bar in ( A ), 1 mm. Scale bar in ( B ), 100 µm. Scale bar in ( C ), 100 µm.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_6833/pmc12726833/pmc12726833__elife-107018-fig2.jpg)