Fig. 1: Kidney cancer progression in the injured kidney is enhanced with an increase in F4/80^lowLy6C^low M2-like macrophages and decrease in tumor-infiltrating T cells.

a uIRI was conducted for 30 min, and mice were euthanized 14 days after uIRI. Kidney fibrosis and macrophage accumulation in the renal cortex was assessed. Representative Sirius red staining (left) and immunohistochemical analysis of F4/80 expression (right) in the kidney cortex are shown. Scale bars: 100 μm. b–g RenCa cells were inoculated into subcapsular kidney 14 days after uIRI (uIRI-Can) or sham-operation (Sham-Can). Mice were euthanized 20 days after inoculation. b Picture of uIRI-Can and Sham-Can is shown. c The tumor volume of uIRI-Can and Sham-Can is shown. (n = 6 per group; two-sided Mann–Whitney U test). d F4/80^lowLy6C^low cells, F4/80^highLy6C^low cells, and F4/80^lowLy6C^high cells were detected by flow cytometry. One plot from each group (Sham-Can (left) and uIRI-Can (right)) is shown. Percentages of F4/80^lowLy6C^low cells (e), F4/80^highLy6C^low cells (f), and F4/80^lowLy6C^high cells (g) among F4/80⁺ cells are shown. (n = 6 per group; two-sided Mann–Whitney U test). h RenCa cells were subcutaneously injected 14 days after uIRI or sham operation, and mice were euthanized on day 20 after tumor injection. The tumor volume of each group is shown. (n = 6 per group; two-sided Mann–Whitney U test). i, j RenCa cells were co-cultured with F4/80^lowLy6C^low or F4/80^highLy6C^low macrophages in Transwell systems for 24 h after creating a gap in the confluent monolayer of RenCa cells, and the degree of gap closure was evaluated. (n = 6 per group). i Representative picture of each group is shown at time 0 and 24 h after starting co-culture. Scale bars: 300 μm. j The quantification of the percentage of the closure area is shown. (two-sided Mann–Whitney U test). k CD3⁺, CD4⁺, and CD8⁺ tumor-infiltrating T cells were detected by flow cytometry. One plot from each group (Sham-Can (left) and uIRI-Can (right)) is shown. l–n CD3⁺, CD4⁺, and CD8⁺ tumor-infiltrating T cell proportion in CD45⁺ cells were quantitatively evaluated in uIRI-Can and Sham-Can, as determined by flow cytometer. (n = 6 per group; two-sided Mann–Whitney U test). o Representative immunohistochemistry images of uIRI-Can and Sham-Can stained for CD3 (left) and CD8 (right) are shown. Scales bar, 100 μm. uIRI unilateral ischemia-reperfusion injury, uIRI-Can kidney cancer inoculated into the kidney subcapsule after unilateral IRI, Sham-Can kidney cancer inoculated into the kidney subcapsule after sham operation.