Figure 6: Active Wnt/β-catenin signaling pathway is necessary for KRT14^pos cell proliferation and bladder repair upon injury.

(a) In vivo treatment with 250 mg kg⁻¹ indomethacin reduces CPP-induced proliferation (8.47±2.05% from 35.74±7.3% in vehicle-treated controls) and KRT14^pos cell numbers (24 h after injury; 10.04±1.45% from 20.17±0.43% in vehicle-treated controls). Data are means of percentages ±s.e.m from four vehicle, or five Indomethacin-treated mice. Total cells counted were 5,108 and 9,947, respectively. (b) Clonogenic assays on Matrigel of total bladder populations cultured in the presence (421.6±10.8) or absence (705±48.4) of 100 μM indomethacin. Data are averages of five independent cultures. (c) Bladder explant cultures with increasing concentrations of indomethacin. (d) Axin 2 transcript levels (quantitative PCR data) in 3- day explants treated with 50 (56.1±2.7%) and 200 μM indomethacin (39.3±6.9%) plotted as percentage of Axin2 levels from vehicle-treated explants. (e) Clonogenic assays on Matrigel of bladder populations (10,000 cells) stably transduced with lentiviral vectors expressing a scrambled (167.38±7.14, n=8) or an anti-β-catenin (β-cat) shRNA (96.86±9.49, n=7). (f) Expression levels of β-catenin in 2-week total Matrigel cultures of primary bladder cells expressing anti-β-catenin (β-cat) shRNA (10.86±1.48%) plotted as percentage of β-cat levels from Matrigel cultures of primary bladder cells expressing scrambled shRNA. All data are mean values±s.e.m. For a, b and e, two group comparison was performed using the Student’s t-test and *P<0.05; **P<0.01. Stars designate the bladder lumen.