Fig. 3

UCHL3 activates the Wnt signaling pathway. A Heatmap showing the significantly differentially expressed genes in UCHL3-deficient T24 cells and wild-type T24 cells. B Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis was conducted to assess the different signaling pathways between wild-type cells and UCHL3-knockout cells. C, D UCHL3 increases the transcriptional activity of the Wnt signaling pathway. Wnt reporter firefly luciferase plasmid (200 ng), CMV (10 ng) and the indicated amounts of UCHL3 plasmid were transfected into 293 T cells. Reporter assays were performed 48 h after transfection, and the results are presented as Wnt/CMV luciferase activity. E Luciferase assays were performed to reveal the influence of UCHL3 loss on the Wnt pathway in T24 cells. F UCHL3 loss inhibited the transcription of Wnt-targeted genes in T24 cells. The expression levels of the indicated Wnt-targeted genes were examined by RT‒PCR in UCHL3^−/− T24 cells and wild-type T24 cells. **P < 0.01; ***P < 0.001; ****P < 0.0001 compared with controls