Fig. 6. Transcriptional factors involved in the Vav2^Onc-dependent transcriptome.

a Transcription factor binding sites found enriched in the promoter regions of up- (red) and downregulated (blue) genes present in the “redundant” fraction of the Vav2^Onc-driven transcriptome. The NES and percentage of hits for each transcription factor binding site are also indicated. b Circos plot of the differentially expressed genes in the Vav2^Onc mouse skin. For each gene, the expression heatmap, the chromosomal location (1–19, X, Y), the binding sites for the indicated transcription factors, and the mirror-image behavior in the Vav2^Onc-regulated transcriptome and the Vav2;Vav3-dependent fraction of the TPA-stimulated gene expression program in the skin of mice are indicated. Red, redundant; Nonred, nonreduntant. c Expression of indicated proteins (left) in the epidermis of the back skin of WT and Vav2^Onc/Onc mice (left), 3D organotypic cultures from WT and Vav2^Onc/Onc primary keratinocytes (middle), and 3D cultures of human keratinocytes (right). In the case of the immunofluorescence experiments using c-Myc antibodies (back skin, green color), sections were decorated with K14 (red color) and DAPI (blue color). Scale bar, 10 μm (n = 3 independent analyses for each experimental group). d Expression of indicated proteins (left) in 3D cultures of control and Rac1^F28L + RhoA^F30L-expressing human keratinocytes. Scale bar, 10 μm (n = 3 independent cultures). e–h Transiently transfected Vav2^Onc triggers the rapid activation of endogenous AP1 (e), c-MYC (f), E2F- (g), and TEAD (h) proteins in human keratinocytes. Activity was measured using luciferase-encoding vectors containing promoter regions for each of the interrogated transcriptional factors, as described in Methods. Data represent the mean ± SEM. Black and gray stars indicate the P value of the indicated experimental values when compared to EGFP- and EGFP-Vav2^Onc-transfected cells, respectively. *P = 0.011 (Vav2^Onc+E200A) versus Vav2^Onc, TEAD); **P = 0.008 (Vav2^Onc versus EGFP, AP1), 0.010 (Vav2^Onc+E200A) versus Vav2^Onc, AP1), 0.002 (Vav2^Onc versus EGFP, E2F); ***P < 0.0001 (all other tests) (ANOVA and Tukey’s HSD test, n = 3 independent experiments). i Expression of indicated proteins in one of the experiments performed in e to h. Tubulin was used as loading control (bottom panel) (n = 3 independent experiments). j, k Correlation between the levels of the VAV2 mRNA and the expression of c-MYC- (j) and YAP/TEAD-regulated- (k) gene signatures in the indicated (top) cSCC (n = 40) and hnSCC (n = 685) gene expression datasets. **P = 0.007 (VAV2 medium, MYC signature, GSE30784), 0.005 (VAV2 high, MYC signature, HPV^- TCGA), 0.008 (VAV2 high, YAP signature, GSE45216), 0.002 (VAV2 medium, YAP signature, GSE30784); ***, P < 0.0001 (all other experiments) using the ANOVA and Dunnett’s multiple comparison test). Data represent the mean ± SEM. Source data for this figure are provided as a Source Data file.