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. 2023 Jan 30;14:489. doi: 10.1038/s41467-023-36008-y

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© The Author(s) 2023

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. 5 — a Distribution of m⁶A peaks across 5′-UTR, CDS, and 3′-UTR of mRNA at day 14 after PT. b Venn diagram showing numbers of genes with significant changes in expression (up: fold change ≥ 2, P < 0.05; down: fold change ≤ 0.5, P < 0.05, rescaled hypergeometric test). c The m⁶A level of Plpp3 transcript was regulated in PT mice after EV-circSCMH1 administration. d, e Effect of EV-circSCMH1 on Plpp3 mRNA (d) and LPP3 (e) levels in mice at day 14 after PT. n = 6 mice/group. Three representative immunoblots were presented from 6 mice/group. ^***P < 0.0001 (d), ^***P = 0.0003 (e) versus sham; ^##P = 0.0030 (e), ^###P < 0.0001 (d) versus PT + EV-Vector. f, g Effect of circSCMH1 plasmid on the expression of Plpp3 mRNA (f) and LPP3 (g) in primary mouse brain microvascular ECs after OGD. Data were presented by three independent experiments. *P = 0.0171 (g), ^***P < 0.0001 (f) versus Con + Vector; ^#P = 0.0375 (g), ^###P = 0.0003 (f) versus OGD + Vector. h Specific primers against m⁶A peak were designed to amplify m⁶A peak of Plpp3 transcript in RNA from the peri-infarct cortex of mice at day 14 after PT. n = 6/each group. ^***P < 0.0001 versus the sham; ^###P < 0.0001 versus the PT + EV-Vector. i Specific primers against the m⁶A peak were designed to amplify the m⁶A peak of Plpp3 transcript in bEnd.3 cells. Data were presented by three independent experiments. ^***P = 0.0002 versus Con + Vector; ^###P = 0.0007 versus OGD + Vector. j, k The bEnd.3 cells were transfected with LV-circSCMH1 and shFTO. The Plpp3 mRNA (j) and LPP3 (k) was detected at 12 h after OGD. Data were presented by 4 (j) or 3 (k) independent experiments. **P = 0.0020 (k), ^***P = 0.0003 (j) versus Con + LV-Vector + shCon; ^##P = 0.0012 (j), ^##P = 0.0087 (k) versus OGD + LV-Vector + shCon; ^†P = 0.0408 (k), ^††P = 0.0083 (j) versus OGD + LV-circSCMH1 + shCon. The data in d, e, h, j, k were expressed as mean ± SEM; one-way ANOVA followed by Holm–Sidak post hoc multiple comparison test. The data in f, g, i were expressed as mean ± SEM; two-way ANOVA followed by Bonferroni’s post hoc multiple comparison tests. Source data are provided as a Source Data file. CDS coding region, shRNA short hairpin RNA, 3′ UTR 3′ untranslated regions, 5′ UTR 5′ untranslated regions.