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. 2020 Jan 31;11:630. doi: 10.1038/s41467-020-14466-y

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© The Author(s) 2020

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. 3 — a Microarray gene expression profile of NSPCs treated for 12 h with fibrinogen compared to control cells. Heatmap analysis showing genes regulated by a factor of at least 4 between fibrinogen-treated and control NSPCs. (n = 3). b Immunolabeling for phosphorylated Smad1/5/8 (P-Smad1/5/8, red) in fibrinogen-treated hippocampal NSPCs. White boxes indicate enlargement of nuclear P-Smad1/5/8- and P-Smad1/5/8 + cell in untreated and fibrinogen-treated conditions, respectively. Scale bar, 46 μm. Quantification of nuclear P-Smad1/5/8. (n = 2, experiments performed in duplicates (mean ± s.e.m., unpaired Student’s t-test, ***P < 0.001). c Immunoblot for P-Smad1/5/8 and Smad1 in NSPCs pretreated with a BMP receptor inhibitor before fibrinogen stimulation. BMP-2 served as positive control. d GFAP + astrocytes in NSPC cultures pretreated with a BMP receptor inhibitor 1 h before fibrinogen treatment 2 days after initiation of differentiation. Scale bar, 125 μm. Quantification of GFAP + astrocytes. (n = 5, (mean ± s.e.m, one-way ANOVA and Bonferroni’s multiple comparisons test, **P < 0.01, ***P < 0.001, ****P < 0.0001). e GFAP + astrocytes (green) in fibrinogen-treated Id3^−/− and WT NSPCs cultures after 2 days on poly‐D‐lysine. Scale bar, 72 μm. Quantification of GFAP + astrocytes. (n = 4, WT; n = 3, Id3^−/− cells, mean ± s.e.m, unpaired Student’s t-test, *P < 0.05). f Experimental setup for photothrombosis on Nestin-CreER^T2::Rosa26-yfp mice. TAM: tamoxifen (right, top). Id3 (red) and YFP (green) immunostainings in the SVZ of uninjured mice and of ancrod-treated mice compared to control WT mice 1 day after PT. The white boxes indicate the enlargement of an Id3 + YFP + (right, top) and an Id3-YFP + (right, bottom) cell in the SVZ of control mice and fibrinogen-depleted mice, respectively, 1 day after PT. Scale bars, 30 μm, left and 8 µm, enlargement. Quantification of Id3 + YFP + cells in the SVZ per area. (n = 4 uninjured mice, n = 4 control mice after PT, n = 3 ancrod mice after PT, unpaired Student’s t-test, *P < 0.05).