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. 2022 Apr 22;13(4):397. doi: 10.1038/s41419-022-04854-1

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© The Author(s) 2022

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. 4 — A–D tube formation assay. A, C Representative images and (B, D) quantitative analysis of tube formation assay characterizing total master segment length, junction numbers, and mesh numbers in H1299 cells (A, B) or SCC9 cells (C, D) transfected with shRNAs for p63 or/and Ago2 as indicated. n = 3 per group. E–H CAM assays. E, G Representative images and (F, H) quantitative analysis of the CAM assay characterizing neovascularization in the chick CAM of H1299 clones (E, F) or SCC9 clones (G, H) expressing shRNAs for p63 or/and Ago2 as indicated. H1299 clones (5 × 10⁶ cells) or SCC9 clones (2 × 10⁷ cells) stably expressing shRNAs for p63 and/or Ago-2 were suspended in Matrigel and then inoculated onto the chick CAM. After 11 days, the primary tumors were imaged at 2X by a dissecting microscope. Images are representative of tumors from three separate experiments. Magnified images indicated by the arrow represent the typical vascular area (size: 1.5 × 1 inches in the original image (7.2 × 5.6 inches)) on tumors. Neovascularization was quantified by counting the number of capillaries of the typical vascular area (1.5 × 1 inches) on the tumor. All experiments were performed in triplicate. *p < 0.05; **p < 0.01; ***p < 0.001.