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. 2023 Aug 4;14(8):500. doi: 10.1038/s41419-023-05995-7

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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. 6 — A Number of primary neurospheres obtained from the hippocampus of wild-type (N = 7), Kidins220^f/f (N = 6), and Kidins220^GfapΔ/Δ (N = 10) mice. B Diameter of wild-type (N = 25), Kidins220^f/f (N = 19), and Kidins220^GfapΔ/Δ (N = 26) primary hippocampal neurospheres from N = 3 mice from each genotype. C Representative phase contrast images of primary hippocampal neurospheres from wild-type, Kidins220^f/f, and Kidins220^GfapΔ/Δ mice. Scale bar, 50 μm. D Number of primary neurospheres obtained from the walls of the lateral ventricles of wild-type (N = 12), Kidins220^f/f (N = 12), and Kidins220^GfapΔ/Δ (N = 15) mice. E, mean neurosphere diameter of SEZ neurospheres of wild-type (N = 9), Kidins220^f/f (N = 7) and Kidins220^GfapΔ/Δ (N = 8) mice. F, Cumulative number of cells obtained over 3 consecutive passages of SEZ neurospheres (N = 7 independent cultures per genotype). G Percentage of Ki67⁺ cells in SEZ neurospheres of the three genotypes (wild-type, N = 3; Kidins220^f/f, N = 2; Kidins220^GfapΔ/Δ, N = 3). H Representative confocal micrographs of wild-type and Kidins220^GfapΔ/Δ SEZ neurospheres stained for the cell cycle marker Ki67 (red) and DAPI (blue) to counterstain the nuclei. Scale bar: 10 μm. I Quantitative analysis of the percentage of live cells (Annexin V^-/7AAD^-) and J, of early-stage apoptotic cells (Annexin V⁺/7AAD^-) in wild-type (N = 14), Kidins220^f/f (N = 6) and Kidins220^GfapΔ/Δ (N = 11) NSCs from the SEZ. K Representative FACS dot-plots of SEZ NSC double-stained for Annexin V/7-AAD. L, Quantification of the number of SEZ neurospheres from Kidins220^GfapΔ/Δ formed after a treatment with 25 μM Z-VAD to inhibit caspases or vehicle for 4 days (N = 4 independent experiments). M Representative phase contrast micrographs of Kidins220^GfapΔ/Δ SEZ neurospheres in both conditions, scale bar: 50 μm. Data represent mean ± s.e.m. Each data point represents values from a neurosphere culture established from an individual mouse (except in B), or an individual mouse (O). Ns, not significant, *P < 0.05, **P < 0.01, ***P < 0.001, by two-tailed paired t test (L), one-way ANOVA (A, B, D, E, G, I, J) and two-way RM ANOVA (F) followed by Dunnett’s post-hoc tests. 7AAD, 7-aminoactinomycin D; FACS, fluorescence-activated cell sorting.