Fig. 6: YAP promotes mTOR signaling.

a GSEA for TEAD, YAP, and mTOR signatures. See Supplementary Data 3 for gene set details. b Representative IHC and c quantification of phospho-S6-positive cells in basal epithelial cells (N = 3, E = 4, Y = 3, EY = 4 mice); scale bars: 20μm (top), 5μm (bottom). d phospho- and total EGFR and S6 expression in Cal27 whole cell lysate following siRNA-mediated knockdown of YAP, TAZ, and YAP + TAZ. e Nrg1 expression in epithelia by RNAseq (N = 5, E = 5, Y = 3, EY = 6 mice). f YAP CUT&Tag, H3K27ac CUT&Tag, and ATACseq peaks at Nrg1 intron 1. Full Nrg1 locus shown in Supplementary Fig. 9. g NRG1, EREG, EPGN on phospho- and total HER3, and S6 expression in Cal27 lysate following siRNA-mediated knockdown. h Axl expression in transgenic tongue epithelia by RNAseq (N = 5, E = 5, Y = 3, EY = 6 mice). i YAP CUT&Tag, H3K27ac CUT&Tag, ATACseq peaks at Axl. j Left: Immunoblots of Cal27 lysate. Right: AXL immunoprecipitation followed by immunoblot for phospho-tyrosine (pY) and AXL following treatment with AXL (R428), EGFR (Erlotinib), and mTOR (INK128) inhibitors. k Representative wells (top) and quantification (bottom) of clonogenic assays in Cal27 treated with DMSO, R428, INK128, or Erlotinib (Erlo) (n = 4 mice per group). l Proposed model for YAP-mediated activation of mTOR via AXL, NRG1, and the EGFR/HER3 axis. Created in BioRender. https://BioRender.com/s33u628. m In vivo mTOR inhibition with rapamycin. Created in BioRender. https://BioRender.com/y80u152. Carcinoma (n) frequency and (o) burden after treatment of transgene-induced EY mice with control (n = 23) or rapamycin (n = 11). Panel n: Fisher’s exact test, two-sided p-values; Panel o two-tailed Mann-Whitney test. p Representative H&E staining of n = 23 vehicle- and n = 11 rapamycin-treated EY mouse tongues. Scale bars both 100 μm. Panels a, c, e, h, and k: ANOVA with Tukey correction for multiple comparisons. All panels: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Panels c, e, h, k, and o display biological replicates. Boxplots in c, e, h, k, and o: median, IQR, and range (or median and range for conditions with 3 points). Source data provided for c, e, h, k, n, and o. For f, i black bars indicate significant peaks, red bars indicate EY-gained peaks.