Fig. 2: Upregulation of Cxcl12 expression in p16^h-sn fibroblasts from the aged bladder.

a, Uniform Manifold Approximation and Projection (UMAP) visualization of total Tomato⁻ (p16^l) and Tomato⁺ (p16^h-sn) bladder cells from an aged (25-month-old) male p16-Cre^ERT2-tdTomato mouse, colored by clusters. b, Distributions of Tomato⁻ and Tomato⁺ cells in the UMAP. c, Numbers of Tomato⁻ (p16^l) and Tomato⁺ (p16^h-sn) cells among the indicated cell types. d, Cell type composition of Tomato⁺ (p16^h-sn) cells. e, Volcano plot showing DEGs between Tomato⁺ (p16^h-sn) and Tomato⁻ (p16^l) fibroblasts from the bladder. The red and blue dots indicate upregulated and downregulated DEGs, respectively, identified by an adjusted P value of <0.05 and a log₂ fold change (log₂FC) value of >0.15. This analysis was two-sided, and Benjamini–Hochberg adjustment was performed. f, Representative immunofluorescence image of bladder fibroblasts from young (7- to 12-week-old) male p16-Cre^ERT2-tdTomato mice, using the indicated staining. White arrowheads indicate double-positive cells for both Tomato and CXCL12. Scale bar, 200 μm. g, Percentages of CXCL12⁺ cells in Tomato⁻ (p16^l) and Tomato⁺ (p16^h-sn) fibroblasts (n = 3 for each group). h, Top 15 enriched GO terms derived from upregulated DEGs of Tomato⁺ (p16^h-sn) fibroblasts. Red boxes indicate GO terms related to cancer progression. GAG, glycosaminoglycan. All terms were identified by an adjusted P value of <0.05. A paired t test was performed for g. Data in g are presented as mean ± s.e.m. All t tests were two-sided.

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