Extended Data Figure 9 : Functional and molecular characterization of skin papillomas after Sox2 deletion.

From: SOX2 controls tumour initiation and cancer stem-cell functions in squamous-cell carcinoma

Extended Data Figure 9

a, Genetic strategy used to study the role of SOX2 in pre-established skin tumours. b, Protocol of DMBA/TPA and tamoxifen (TAM) administration. c, Macroscopic pictures of skin papillomas after Sox2 deletion. Tamoxifen administration to K14CreER:SOX2fl/fl mice presenting with skin papillomas leads to their regression. d, Co-immunostaining for K14 (green) and SOX2 protein (red) in control (Ctrl) and Sox2 conditional knockout (cKO) papilloma after 1 week of tamoxifen administration showing the disappearance of SOX2 expression in the tamoxifen-treated conditions. e, Co-immunostaining for K14 (green) and K10 (red) in control and Sox2 conditional knockout papilloma after 1 week of tamoxifen administration showing the decrease in the number of differentiated K10+ cells in Sox2 conditional knockout tumours. f, Co-immunostaining for K14 (white), Pdpn (red) and SOX2 protein (green) in control and Sox2 conditional knockout papilloma after 1 week of tamoxifen administration. g, Histograms summarizing the genes downregulated in Sox2 conditional knockout TECs of papillomas (n = 3 microarrays for each group and the histograms show the mean and s.e.m.). These data show that SOX2 controls a gene network that regulates tumour stemness, proliferation/survival, metabolism, cell adhesion/invasion, transcription and chromatin remodelling factors, and paraneoplastic hypercalcaemia. h, Venn diagram showing the overlap between the genes upregulated in the SOX2+ CSC signature and downregulated following Sox2 deletion. The arrow indicates the hypergeometric P value of this overlap. These data show that genes preferentially expressed by SOX2–GFP+ CSCs and positively controlled by SOX2 are significantly enriched. Genes of this overlap are presented in Supplementary Table 1. i, Venn diagram showing the overlap between the genes downregulated following SOX2 deletion in skin TECs and downregulated in inducible Sox2-null mouse embryonic stem cells (Sox2 cKO)27. The 57 genes of the overlap and the 46 genes downregulated in the SOX2-regulated gene signature and bound by SOX2 in embryonic stem cells28 are presented in Supplementary Table 2. j, Venn diagram showing the overlap between the genes downregulated in the SOX2-regulated gene signature and genes downregulated (fold change >2) in a transient knockdown of SOX2 in a human glioblastoma cell line (SOX2 KD)29. The 5 genes of the overlap and the 49 genes downregulated in the SOX2-regulated gene signature and bound by SOX2 in the human glioblastoma cell line are presented in Supplementary Table 3. Scale bars, 50μm. Down, downregulated genes; Epi, epithelium; ESC, embryonic stem cells; GB, glioblastoma cell line; Str, stroma; Up, upregulated genes.