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. 2023 Feb 8;20:28. doi: 10.1186/s12974-023-02699-9

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© The Author(s) 2023

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 5 — CD47-deficient RPE cells lose melanosomes/melanolipofuscin to melanophages. Gating and FarRed CellTrace intensity measurements by cytometry of human CD14 + Mo after 2 h of incubation with a monolayer of FarRed CellTrace pre-stained ARPE19 cells (a human RPE cell line) with 10 µg/ml of a control antibody (black line) or CD47 blocking antibody B6H12 (red line A) and quantification of the fluorescence intensity (B; n = 6 wells per group from three independent experiments; Mann–Whitney p = 0.0221). Three independent experiments gave similar results. Representative micrographs of phalloidin (red fluorescence staining), IBA1 (green fluorescence staining) double-labeled RPE/choroidal flatmounts of 12-month-old Cd47^+/+ (upper panel) or Cd47^−/−- recipient mice (lower panel) that had received a Cd47^+/+- bone marrow transplant after lethal irradiation at 6 months of age (C). Quantification of the number of IBA1-stained subretinal MPs (D) and quantification of the percentage of pigment-laden melanophages (that block the phalloidin staining of the underlying RPE) of total subretinal MPs (E) of 12-month-old WT and Cd47^−/−-mice compared with Cd47^+/+ bone marrow transplanted WT and Cd47^−/−-mice (n = 5/group; Mann–Whitney p = 0.00,159). Scale bar = 20 µm