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. 2021 Oct 18;12:6072. doi: 10.1038/s41467-021-26346-0

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 8 — (A, quantified in B) Intra-peritoneal (IP) treatment with Hepta-ANGPT1 from birth to P14 led to increased SC area in WT (p = 0.0014) and (C, quantified in D) Angpt1ΔNC eyes (p = 0.0008). Wildtype, n = 4 (vehicle) and 10 (Hepta-ANGPT1), Angpt1ΔNC, n = 4 (vehicle) and 8 (Hepta-ANGPT1) animals from four independent litters. PROX1 expression in WT eyes was also increased, suggesting that this larger SC retained its fully differentiated phenotype (p = 0.0109). E Hepta-ANGPT1 treatment had no effect on the morphology of the limbal superficial capillary plexus suggesting the effect was specific to Schlemm’s canal. F Some Angpt1ΔNC eyes treated with Hepta-ANGPT1 exhibited a sprouting endothelial plexus in place of an organized, canal-like structure. G Intraocular pressure was measured at 8, 10, and 15 weeks in mice that received Hepta-ANGPT1 from birth to P14. Wildtype n = 12 (vehicle) and 13 (Hepta-ANGPT1), Angpt1ΔNC n = 10 (vehicle) and 14 (Hepta-ANGPT1) from four independent litters. H Following IOP measurement, average Schlemm’s canal area was measured in whole mount eyes by confocal microscopy and I BRN3A-positive retinal ganglion cells were quantified in retina flat mounts of the same animals. J To examine the acute effect of Hepta-ANGPT1 on IOP in adult WT eyes, 8-week-old C57Bl/6J mice received bilateral intravitreal injections of either 1 μg Hepta-ANGPT1 or PBS vehicle after 3 days of baseline IOP measurement. IOP was then measured daily for 5 days following injection. n = 12 (vehicle) and 13 (Hepta-ANGPT1). Scale bars represent 100 μm, quantified fields represent a total area of 262,277 μm² (B, D, H) and 65,413 μm² (I). AFU, background-subtracted arbitrary fluorescence units. Dashed lines in PROX1 panels indicate CD31 + SC area. *p < 0.05, **p < 0.01, ***p < 0.001 as determined by two-tailed Welch’s t-test (B, D) Df = 9 (WT, B) and 8 (Angpt1ΔNC, D) or two-way ANOVA followed by Bonferroni’s correction (G–J). Df_group = 3, F_group = 172.2 (G), Df_treatment = 1, F_treatment = 4.6 (H), Df_group = 3, F_group = 11.92 (H), Df_treatment = 1, F_treatment = 24.03 (J).