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. 2023 May 19;14:1197709. doi: 10.3389/fimmu.2023.1197709

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Copyright © 2023 Seiler, Kähler, Kleingarn, Sadik, Bieber, Köhl, Ludwig and Karsten

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Characterization of the cell-specific C5aR2 knock-out mouse line. (A) PCR-based detection of the floxed tdTomato-C5ar2 gene cassette in BM neutrophils (N), BM Ly6G^-/CD11b⁺ cells (C), splenic B (B) and T cells (T) from LysM ^cre -C5ar2 ^–/– mice. Positive control from C5ar2-tdTomato ^fl/fl mice (PC) and No Template Control (NTC) are shown. M: GeneRuler 50 bp DNA Ladder. (B) Histogram overlay of tdTomato reporter signal indicating C5aR2 expression in BM and blood neutrophils (Ly6G⁺/CD11b⁺) and Ly6G^-/CD11b⁺ cells from LysM ^cre -C5ar2 ^–/– (purple) and C5ar2-tdTomato ^fl/fl (red) mice. The tdTomato signal of corresponding cells from WT (grey) mice served as a negative control. (C) Histogram overlay indicating C5aR1 expression in BM and blood neutrophils from LysM ^cre -C5ar2 ^–/– (purple), C5ar2-tdTomato ^fl/fl (red), and WT (grey) mice. The signal corresponding to the FMO control is shown in black. (D) C5aR1 expression in BM and blood neutrophils (Ly6G⁺ cells) from LysM ^cre -C5ar2 ^–/–, C5ar2-tdTomato ^fl/fl, and WT mice quantified by the gMFI. (E) Frequency (of parent) of neutrophil subsets in the BM and blood of naïve LysM ^cre -C5ar2 ^–/– and C5ar2-tdTomato ^fl/fl control mice (n = 6/group). * p < 0.05; ns, not significant.