Figure 1. The Trx1 system is largely dispensable for the development and homeostatic maintenance of myeloid cells.

(A–E) Txnrd1^fl/fl;Rosa26-CreERT2 mice and Txnrd1^fl/fl littermates were injected with TAM to delete the Txnrd1 gene and were analyzed by flow cytometry 2 weeks later. Depicted are the total numbers or percentages of the indicated populations in the thymus (A), bone marrow (BM; B), blood (C), lungs (D), spleen (E); n = 4–5 mice). (F–I) Lethally irradiated WT mice (CD45.1⁺CD45.2⁺) were reconstituted with a 1:1 mixture of WT (CD45.1⁺) and TAM-treated Txnrd1^fl/fl;Rosa26-CreERT2 (CD45.2⁺) bone marrows (or Txnrd1^fl/fl as control). After reconstitution, the percentage of donor CD45.2⁺ cells among the indicated cell populations was determined by flow cytometry (n = 4–5 mice). (F) Schematic showing the experimental setup. (G) Depicted are the CD45.2⁺ percentages of total TCRβ⁺ T cells in the indicated organs. (H, I) The percentages of CD45.2⁺ cells among the indicated myeloid populations in the bone marrow (BM; H) and lungs (I) are shown. DP, CD4⁺CD8⁺ double positive thymocytes; CD4⁺ T, CD4⁺ single positive thymocytes; CD8⁺ T, CD8⁺ single positive thymocytes; Eo, eosinophils; Neutro, neutrophils; Mono, monocytes; DC, dendritic cells; AlvMac, alveolar macrophages; cDC1/2, type 1/2 conventional dendritic cells; Mac, macrophages; LN, lymph nodes. Bar graphs show mean + standard deviation (A–E, G–I). Data are representative of three independent experiments. For each panel, a representative experiment with biological replicates (A–E,G–I) is shown. Student’s t test (two-tailed, unpaired) was used for the comparison of two groups (A–E, G–I): *, p≤0.05; **, p≤0.01; ***, p≤0.001; ****, p≤0.0001; ns, not significant.

Figure 1—figure supplement 1. Efficiency of Txnrd1 gene deletion.

(A, B) Txnrd1^fl/fl;Rosa26-CreERT2 mice and Txnrd1^fl/fl littermates were injected with TAM to delete the Txnrd1 gene and were analyzed by flow cytometry 2 weeks later. Depicted is the analysis of genomic Txnrd1 DNA (A) and Txnrd1 mRNA (B) in total bone marrow (BM) cells (left) and in MACS-enriched CD11b⁺ cells from the spleen (right) determined by RT-PCR (n = 3 mice). Bar graphs show mean + standard deviation. Data are representative of two independent experiments. For each panel, a representative experiment with biological replicates is shown. Student’s t test (two-tailed, unpaired) was used for the comparison of two groups: *, p≤0.05; **, p≤0.01; ***, p≤0.001; ****, p≤0.0001; ns, not significant.

Figure 1—figure supplement 2. Gating strategies for the analysis of distinct myeloid-cell populations in vivo.

(A–D) Shown are the gating strategies utilized for the analysis of the indicated myeloid-cell populations in the bone marrow (BM; A), blood (B), lungs (C), and spleen (D). Eo, eosinophils; Neutro, neutrophils; Mono, monocytes; DC, dendritic cells; AlvMac, alveolar macrophages; cDC1/2, type 1/2 conventional dendritic cells; Mac, macrophages.

Figure 1—figure supplement 3. The Trx1 system is largely dispensable for the maintenance and homeostasis of myeloid cells in mixed-bone marrow chimera settings.

(A–C) Lethally irradiated WT mice (CD45.1⁺CD45.2⁺) were reconstituted with a 1:1 mixture of WT (CD45.1⁺) and TAM-treated Txnrd1^fl/fl;Rosa26-CreERT2 (CD45.2⁺) bone marrows (or Txnrd1^fl/fl as control). After reconstitution, the percentage of donor CD45.2⁺ cells among the indicated cell populations was determined by flow cytometry (n = 4–5 mice). (A, B) The percentages of CD45.2⁺ cells among the indicated myeloid populations in the blood (A) and spleen (B) are shown. (C) Depicted are the CD45.2⁺ percentages of total CD19⁺ B cells in the indicated organs. Mono, monocytes; Neutro, neutrophils; Eo, eosinophils; Mac, macrophages; cDC1/2, type 1/2 conventional dendritic cells. Bar graphs show mean + standard deviation. Data are representative of three independent experiments. For each panel, a representative experiment with biological replicates is shown. Student’s t test (two-tailed, unpaired) was used for the comparison of two groups: *, p≤0.05; **, p≤0.01; ***, p≤0.001; ****, p≤0.0001; ns, not significant.