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. 2022 Aug 30;12:14730. doi: 10.1038/s41598-022-19172-x

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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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Ectopic expression of miR-133a strands -3p or -5p inhibits endothelial cell tubular morphogenesis. HUVEC transfected with miRNA “mimics” for strand -3p (miR-133a-3p), -5p (miR-133a-5p), or a negative control (miR-NC) were plated on Growth-Factor Reduced Matrigel (Geltrex) in ECGM medium containing 1% FBS and were left untreated (−) or stimulated with VEGF-A₁₆₅ at 50 ng/ml final concentration (+) as indicated. Images show representative fields from experiments quantified in the histogram. Grey, blue and red bars indicate cells transfected with miR-NC, miR-133a-3p and miR-133a-5p respectively. Data are shown as mean ± SE, n = 7. ns = non-significant; *, P ≤ 0.05; ***, P ≤ 0.001; ****, P ≤ 0.0001 (unpaired two-tailed Student’s t-test when comparing the indicated groups). Scale bars, 1000 µm. ImageJ (https://imagej.nih.gov/ij/) angiogenesis analyzer plugin software was used to quantify “Number of Junctions” in Matrigel assays.