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. 2017 Dec 20;7:17916. doi: 10.1038/s41598-017-17948-0

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© The Author(s) 2017

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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in vivo activity of VHH-PE38 immunotoxins. C57BL/6 mice were injected intravenously with (GFP-specific) Enhancer-, VHH13-, VHH16- or VHH21-PE38 immunotoxins. After 24 hours myeloid cell composition was monitored in the spleen and bone marrow. (A) Dot plot shows FSC-A vs SSC-A parameters of whole bone marrow of mice injected with indicated immunotoxins. (B) CD11b expression on live bone marrow cells. (C) Percentage of CD11b⁺ cells in living (SYTOX Blue negative) bone marrow cells. (D) Ly-6G vs Ly-6C expression gated on CD11b⁺ live (SYTOX Blue negative) bone marrow cells. (E) Percentage of CD11b⁺Ly-6C⁺Ly-6G⁻ and CD11b⁺Ly-6C^lowLy-6G⁺ in living bone marrow cells. (F) CD11b expression on live (SYTOX Blue negative) splenic cells. (G) Percentage of CD11b⁺ cells in live (SYTOX Blue negative) splenic cells. (H) Ly-6G vs Ly-6C expression gated on CD11b⁺ live (SYTOX Blue negative) splenic cells. (I) Percentage of CD11b⁺Ly-6C⁺Ly-6G⁻ and CD11b⁺Ly-6C^lowLy-6G⁺ in live (SYTOX Blue negative) splenic cells. (J) Bar histograms show the CD11b⁺Ly-6C⁺Ly6G⁻/CD11b⁺Ly-6C^lowLy-6G⁺ cell ratio in the bone marrow. (K) Bar histograms show the CD11b⁺Ly-6C⁺Ly6G⁻/CD11b⁺Ly-6C^lowLy-6G⁺ cell ratio in the spleen. *p value < 0.05, **p value < 0.01 on Student-T test. Representative of 2 independent biological experiments.