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. 2020 Oct 20;28(3):1041–1061. doi: 10.1038/s41418-020-00636-4

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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. 1 — A Schematic diagram demonstrating the study design of in vivo experiments on MNU-induced retinal degeneration. B Biodistribution of PKH26-labeled MSCs in the eye after intravitreal injection for 6 h. NC negative control, injection of MSCs without staining. Scale bar = 2.5 mm. C Tracing of PKH26-labeled MSCs (red) in the retina tissue counterstained by Hoechst 33342 (blue) after intravitreal injection for 24 h. MSCT mesenchymal stem cell transplantation after MNU injection, INL inner nuclear layer, ONL outer nuclear layer. Scale bar = 50 μm. Representative hematoxylin and eosin (H&E) staining images of retinal tissues (D) and the corresponding quantitative analysis of ONL thickness (E). Ctrl control, no MNU treatment, GCL ganglion cell layer, ONH optic nerve head. Scale bars = 50 μm. *P < 0.05 by the Kruskal–Wallis test for area under curve (AUC). N = 3 per group. Representative scotopic electroretinography (ERG) waveforms (F) and the corresponding quantitative analyses of amplitude changes of a-wave (G) and b-wave (H). *P < 0.05 by the Kruskal–Wallis tests. N = 6 per group. Representative photopic ERG waveforms (I) and the corresponding quantitative analysis of b-wave amplitude changes (J). *P < 0.05 by the Kruskal–Wallis tests. N = 6 per group. K Representative immunofluorescent (IF) staining images of retinal tissues showing cone photoreceptor bodies (white) counterstained by Hoechst 33342 (blue). Scale bars = 25 μm. Representative terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL, green) staining images of retinal tissues counterstained by Hoechst 33342 (blue) (L) and the corresponding quantitative analysis of percentages of TUNEL⁺ cells over total ONL cells (M). Scale bars = 50 μm. *P < 0.05 by the Kruskal–Wallis test. N = 6 per group. Representative flow cytometric images showing death of 661W cone photoreceptor cells (N) and the corresponding quantitative analysis of percentages of apoptotic (Annexin V⁺PI⁻ plus Annexin V⁺PI⁺) 661W cells (O). The MSC group, Transwell co-culture of MNU-treated 661W cells with MSCs. PI propidium iodide. *P < 0.05 by the Kruskal–Wallis test. N = 3 per group. Data represent median ± range for (E) and (O). Data are represented as box (25th, 50th, and 75th percentiles) and whisker (range) plots for (G), (H), (J), and (M).