Fig. 3

Age associated increase in B cell populations in female mouse bladder. A Healthy male and female mouse bladders (n = 3/4) of ages, 3, 6, 9, 12, 15 and 18 months were subjected to bulk RNA-Seq. Differential expression was assessed using DESeq2 in R (1.4.0) with a P-adj < 0.05 (FDR). Analysis of differential expression gene lists was performed using the MyGeneset application at https://www.immgen.org/, to determine immune cell subset type enrichment. Plot represents comparison between top 200 differentially expressed genes between 15-month-old female and age matched males. B Hematoxylin and eosin stained section showing lymphoid aggregate in an 18-month-old female mouse bladder wall (a). Whole bladder image captured using Phenochart Software image viewer 1.0.9 (Akoya Biosciences) from multiplex immunofluorescence stained 18-month-old female bladder section showing tertiary lymphoid structures (b; white boxes). Immune cells within TLSs (F4/80+ macrophages, CD163+ M2 macrophages, Pax5+ B cells, CD3+ T cells, CD8+ cytotoxic T cells and PD-L1 immune checkpoint (c, d). Representative of n = 3–5 in each sex