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. 2024 Jun 7;4(8):1102–1120. doi: 10.1038/s43587-024-00635-x

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© The Author(s) 2024

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 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/.

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Extended Data Fig. 5 — a. Representative IHC for myeloperoxidase (MPO) from kidneys harvested from 18-month-old Rragc^+/+ and Rragc^S74N/+ male mice. Bars represents 100 μm. b. Representative IHC for myeloperoxidase (MPO) from livers harvested from 18-month-old Rragc^+/+ and Rragc^S74N/+ male mice. Bars represents 100 μm. c. Volcano plot from RNA-seq performed in 6-8-week-old Rragc^S74N/+ (n = 4) versus 6-8-week-old Rragc^+/+ (n = 5) neutrophils isolated from BM. Differential expression was performed with DESeq, using a 0.05 FDR. d. Gene set enrichment analysis (GSEA) related to TFEB target genes, NFKBIA target genes and genes involved in apoptosis in 6-8-week-old Rragc^S74N/+ (n = 4) versus 6-8-week-old Rragc^+/+ (n = 5) neutrophils isolated from BM. e. Quantification of aged neutrophils (CD11b⁺/Ly6G⁺/CD62L^-/CXCR4⁺, left) and fresh neutrophils (CD11b⁺/Ly6G⁺/CD62L⁺/CXCR2⁺, right) by flow cytometry in 2-month-old Rragc^+/+ (n = 2) and Rragc^S74N/+ (n = 3) male mice and in 16-month-old Rragc^+/+ (n = 5) and Rragc^S74N/+ (n = 5) male mice. Statistical significance was calculated by two-way ANOVA. Data are presented as mean values ± SD. f. Experimental setup of the neutrophil adoptive transplant experiment. BM-derived neutrophils were isolated from 10-week-old Rragc^+/+ mice (CD45.1.2) and 10-week-old Rragc^S74N/+ (CD45.2.2) and transplanted at 1:1 ratio to 16-week-old Rragc^+/+ (CD45.1.1) hosts. g. Ratio of CD45.2.2/CD45.1.1 transplanted neutrophils in blood, spleen and BM 16 h (n = 3 biological replicates) and 40 h (n = 3 biological replicates) after adoptive transfer as described in f. Data are presented as mean values ± SD. Statistical significance was calculated by two-tailed Student’s t-test. h. Representative images of IF depicting CD45.1 positive cells in kidneys from young male hosts. Bars represent 20 μm. i. Representative images of IF depicting CD45.1 positive cells in kidneys from old male hosts. Bars represent 20 μm.

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