Defining murine monocyte differentiation into colonic and ileal macrophages
- Weizmann Institute of Science, Israel
Figures
Figure 1 with 4 supplements
Transcriptome analysis of CX3CR1+ monocyte graft - derived colonic and ileal macrophages isolated from macrophage depleted animals.
(A) Experimental protocol. Briefly, [CD11c-DTR > C57BL/6] BM chimeras were treated as indicated with DTx. 2 × 106 CD117- CD11b+ CD115+ Ly6C+ GFPint BM monocytes isolated from Cx3cr1GFP/+ mice were …
Figure 1—figure supplement 1
Comparison of gene expression in monocytes and resident colonic and ileal macrophages.
(A) Sorting schemes for BM Monocytes and resident intestinal macrophages isolated from CX3CR1-heterozygous CX3CR1GFP/+ C57BL/6 mice according to the following surface markers: CD117-CD11b+ CD115+ …
Figure 1—figure supplement 2
Analysis of host tissue for graft-derived cells.
(A) Fluorescent images of ileum of CX3CR1GFP/+ mouse (top) and DTx-treated [CD11c-DTR > WT] BM chimeras engrafted with CX3CR1GFP/+ monocytes at the indicated days post-transfer. (B) Flow cytometric …
Figure 1—figure supplement 3
Pathway analysis of graft-derived cells.
(A) Elbow plot to determine number of clusters. Since the plot was continuous, visual inspection was performed with different cluster numbers; according to cluster homogeneity we selected k = 7. (B) …
Figure 1—figure supplement 4
Analysis of expression of pro-inflammatory genes in adoptive transfer model and DSS-induced colitis model.
Expression of genes in engrafted colonic and ileal macrophages, resident colonic and ileal macrophages at steady state, infiltrating monocytes and resident macrophages isolated from the colon of …
Figure 2 with 2 supplements
Differences between acute monocyte graft -derived macrophages and resident intestinal macrophages.
(A) Volcano plots of genes that change from engrafted macrophages at day 12 and resident macrophages in the colon (left) and ileum (right). Gray dots represent genes that do not significantly change …
Figure 2—figure supplement 1
Comparison of graft and engrafted cells to recent monocytic infiltrates in the colon, identified by their Ly6C surface expression.
(A) Representative flow cytometric analysis of colonic myeloid cells. Recent monocytic infiltrates and macrophages in the colon were defined as CD45+ CD11b+ Ly6C+ MHCII- and CD45+ CD11b+CD64+ Ly6C- …
Figure 2—figure supplement 2
Comparison of colonic macrophages generated upon monocyte transfer into [CD11c-DTR > WT] and [CX3CR1-DTR > WT] BM chimeras treated with DTx.
(A) Venn diagrams of gene lists up- and downregulated as compared to the monocyte graft. (B) Representative upregulated genes similarly expressed between the colonic macrophages as compared to the …
Figure 3
Changes in gene expression during conversion of monocytes into colonic or ileal macrophages.
(A) Example genes that are significantly up-regulated in intestinal macrophages from monocytes to day 4: either up-regulated in the colon but do not change in the ileum, or significantly …
Figure 4
Changes in gene expression during conversion of monocytes into generic intestinal macrophages.
(A) Dot plot for genes whose expression changes at least 2-fold between both monocytes to engrafted colonic macrophages at day four and monocytes to engrafted ileal macrophages at day 4. Black dots: …
Figure 5
Transcription factors which change during intestinal macrophage differentiation.
(A) Expression graphs of 3 TF families: Cebp (left), E2f (middle) and Egr (right). Lines mark average normalized reads, error bars represent SEM. (B) Representative TFs that are down-regulated in …
Figure 6 with 1 supplement
Gene expression changes in Ly6C+ monocytes after differentiating to tissue-resident cells.
(A) Scheme of monocyte development and fates during physiology and inflammation. (B) Fold change heat map of all 1303 genes that significantly change during the transition of Ly6C+ blood monocytes …
Figure 6—figure supplement 1
Gating strategy for sorting of Ly6C+ and Ly6C- blood monocytes.
Representative flow cytometric analysis indicating gates used for isolation of blood monocyte subsets.
Additional files
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Source data 1
Excel sheets of the data which formed the basis of Figures 1–4, and 6.
- https://cdn.elifesciences.org/articles/49998/elife-49998-data1-v1.xlsx
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Supplementary file 1
List of RNAseq data sets prepared and used in this study.
- https://cdn.elifesciences.org/articles/49998/elife-49998-supp1-v1.docx
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Transparent reporting form
- https://cdn.elifesciences.org/articles/49998/elife-49998-transrepform-v1.pdf
