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. 2022 Feb 17;13:945. doi: 10.1038/s41467-022-28593-1

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© The Author(s) 2022, corrected publication 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 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. 4 — A Middle cerebral artery occlusion (MCAO) was induced in C57BL/6 mice for 45 min. Coronal (approx. bregma 0–1 mm) and transversal cryosections were prepared of 24 and 72 hours post-MCAO brains and were co-stained for F4/80 and Osteopontin (Spp1). Overview images of the lesioned striatum (str) and the corpus callosum (cc) and corresponding z-stacks are shown. B The density of Osteopontin⁺-cells was quantified in eight defined regions 1–6 (covering most of the middle cerebral artery flow area at bregma, see numbered rectangles) generated from brains post 24 h (n = 6) and post 72 h (n = 5). Data are presented as mean values ± SEM (two-sided t-test, *p = 0.0106). C Heatmaps were generated from brains post 24 h (n = 6) and post 72 h (n = 5) (two-sided t-test, *p < 0.05). 3D heatmap of Osteopontin expression was constructed by combining nine consecutive transversal sections of 72 h post-MCAO brains (n = 3) using Free-D software. Neuronal-marker MAP2 was used to define lesion borders. D Sections of 24 h and 72 h post-MCAO brains as in A were co-stained for F4/80 together with LPL, M-CSF, and ADAM8. Representative Z-stacks are shown. The density of positive cells was quantified within the defined sections 1–6 of 24 h (n = 6) and 72 h (n = 4) post-MCAO brains. Plots in B and D are representative of n = 4 stainings (sham), n = 6 stainings (24 h post MCAO) and n = 4 stainings (72 h) (two-sided t-test, **p < 0,01, ***p < 0.001; LPL/F4/80: p = 0.0087; M-CSF/F4/80: p = 0.0008; ADAM8/F4/80: p = 0.0043). Data are presented as mean values ± SEM. E Mice (n = 3) were injected with idoxuridine (IdU) twice 48 h and 24 h prior to induction of MCAO and IdU was co-stained with M-CSF. F Sections as in A were co-stained with FluoroMylein and SAMC-marker M-CSF, showing potential myelin phagocyting by CD45- and M-CSF-expressing cells. G M-CSF-expressing cells potentially phagocyting APC⁺-oligodendroglia. H LPL⁺-cell enclosing a NeuN⁺-neuron within the lesioned striatum. I Representative immunofluorescence images and corresponding z-stacks of SAMC-marker Osteopontin, LPL, M-CSF, Adam8, and MMP12 co-stained with microglial/monocyte marker IBA1 within the infarcted tissue of stroke patients (representative images of n = 5 patients). J Human stroke sections stained for SAMC-marker M-CSF and neuronal-marker MAP2 showing potential neuronal phagocyting by M-CSF⁺ cells. Source data for B and D are provided as a Source Data file.