Fig 2: Immunofluorescence for Vim (red) and TRP or Piezo channels (green) in cultured LP-ICs. Double staining reveals the protein expression of TRPV4 (A), piezo1 (B), TRPA1 (C), TRPV2 (D), TRPV1 (E), TRPM8 (F) and Piezo2 (G) in most of the Vim+ LP-ICs. The nucleus marker (DAPI) is stained in blue.

Fig 3: The expression levels of TRPA1 were decreased in the mucosa of human fibrotic bladders. (A) Masson’s staining of a human neurogenic bladder (right) showed significant collagen deposition (blue) in the suburothelial layer when compared to normal bladder (left). Scale bar = 250 µm. (B) Immunohistochemical analysis of a-SMA in the suburothelial layer indicating a significant increase in a-SMA expression in fibrotic bladder. Scale bar = 100 µm. The red arrows indicate positive staining for a-SMA. (C) Immunofluorescence analysis of TRPA1 (green) and vimentin (Vim, red) in the suburothelial layer showing reduced expression of TRPA1 in Vim-expressing cells in the submucosa of fibrotic human bladder when compared to normal human bladder. Scale bar = 20 µm. (D) Analysis of the mRNA levels of TRPA1, a-SMA, COL1A1, COLIII, FN, and TGF-ß1 in mucosa, indicating a significant increase in the expression of a-SMA, COL1A1, COLIII, FN, and TGF-ß1, but a significant decrease in the expression of TRPA1 in fibrotic bladder when compared with normal bladder. Data are shown as means ± SEM (n = 3 bladders in each group). *** p < 0.001.