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. 2022 Sep 14;13:5376. doi: 10.1038/s41467-022-33117-y

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© This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 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. 5 — a Numbers ± SEM of CD19⁺ (P = 0.0002, left) and CD93⁺CD19⁺ (P = 0.0052, middle) B cells and frequency ± SEM of CSF1R⁺ within CD93⁺CD19⁺ B cells in the spleen of naïve vs 4T1.2 cancer-bearing BALB/c mice (P = 0.0036, right, n = 5). b ELISA measurements of secreted M-CSF in CM by 4T1.2, EMT6, AT3, MC38, B16-F10, and ID8 cells (n = 6, pg/ml). c–e Representative FACS plots of the F4/80⁺CD11b⁺ B-MF converted from highly FACS-purified BM CSF1R⁺ and CSF1R⁻ B-cell precursors and splenic CSF1R⁺ and CSF1R⁻ B cells (c), BMBP after 7-day (d), or pre-B-cell line 70z/3 after 30-day (e) culture in 4T1.2-CM alone or in the presence of neutralizing anti-M-CSF Ab or a specific CSF1R inhibitor Ki20227 (d, e). f, g Representative FACS plot (f) and quantification (frequency ± SEM, g) of CSF1R⁺ BM B cells in Mb1-CSF1R^Flox/Flox mice as compared to WT littermates (P = 0.0002, top panel) used for the generation of B-MF after 7-day 4T1.2-CM treatment (P < 0.0001, lower panel, n = 6 mice). Gating is shown in Supplementary Fig. 8. In a, b, and g, each symbol is for a single mouse. P-values in a, b, and g were calculated using two-tailed unpaired t-test. Results for all panels were independently reproduced at least three times.