Fig. 2: NUMB loss is associated with YAP activation and EMT.

a Venn diagrams show the activated transcriptional regulators identified in NUMB-KD vs. Ctr-KD RT4, RT112 and KK47 NIMBC cells (left) and in NUMB^Low vs. NUMB^High cell lines (right). The blue circle indicates the 4 common YAP/TAZ transcriptional regulators in the two paired comparisons. b Hallmark gene sets from MSigDB positively enriched by GSEA in NUMB^Low vs. NUMB^High cell lines and NUMB-KD vs. Ctr-KD RT4, RT112 and KK47 NIMBC cells. NES, Normalized Enriched Score; p, FDR adjusted two-sided permutation test p-value; n = 2. c, d Active YAP/TAZ (c) and EMT (d) gene signature by GSEA of the 3 NUMB^Low vs. 3 NUMB^High cell lines (left panels), and NUMB-KD vs. Ctr-KD RT4, RT112 and KK47 cells (right panels). ES, Enrichment Score; NES, Normalized Enrichment Score; p, two-sided permutation test p-value; n  =  2. e GSEA showing downregulation of an active YAP/TAZ (top) and EMT (bottom) gene signature in the indicated NUMB^High NUMB-KD cell lines treated with 500 nM verteporfin (VP) vs. vehicle for 24 h. ES, Enrichment Score; NES, Normalized Enrichment Score; p, two-sided permutation test p-value; n = 1. f Left, progression-free survival of the UROMOL patient cohort categorized as YAP/TAZ active or inactive by the 22-gene YAP/TAZ signature. HR, hazard ratio (95% confidence interval); p, two-sided log-rank test p-value; n, patient number. Right, box-violin plot showing the EMT score in the YAP/TAZ active and inactive groups. Boxplots are delimited by 25th and 75th percentiles and show the median (horizontal line) and the mean (X). The whiskers span from the smallest and largest data values within a 1.5 interquartile range. ****, p < 0.0001 by two-sided Welch’s t-test. g Left, association between the NUMB^LESS and the 22-gene YAP/TAZ signatures in the UROMOL cohort (n = 535). ****, p < 0.0001, by two-sided Fisher’s exact test. Right, box-violin plot (see legend to Fig. 2f) showing the EMT score in the NUMB^LESS-Like or NUMB^LESS-Not-Like groups. ****, p < 0.0001 by two-sided Welch’s t-test. h Left, Immunofluorescence of YAP and CYR61 expression (red) and DAPI nuclear stain (blue) in high-grade NUMB^High vs. NUMB^Low NMIBC tumors. Bars, 100 μm. Right, quantification of the % of YAP-high vs. -low expressing cells in 4 NUMB^High and 5 NUMB^Low NIMBC, and CYR61 mean cytoplasmic intensity (arbitrary units, A.U.) of 7 NUMB^High and 8 NUMB^Low NIMBC. Values are expressed as mean ± SEM. *, p = 0.038 for YAP, p = 0.035 for CYR61, by two-sided Welch’s t-test for YAP and two-sided two-sample t-test for CYR61. Source data are provided as Source Data file.