Figure 3
From: The pluripotency factor NANOG promotes the formation of squamous cell carcinomas
NANOG promotes epithelial-mesenchymal transition (EMT) in vivo.
(A) Gene set enrichment data (GSEA) showing the enrichment of three published EMT gene signatures23,24,25 in TG papillomas as compared with CTR papillomas. NES: normalized enrichment score. The false discovery rate (FDR; q-value) is indicated. (B) Relative gene expression (fold change) in CTR and TG papillomas and SCCs of EMT related genes analyzed by qRT-PCR. mRNA levels were normalized by Gapdh levels and then expressed as the ratios with respect to CTR papillomas (CTR PAP). (C) Relative mRNA levels of the indicated genes in primary keratinocytes extracted from CTR and TG newborn pups (n = 2) and treated during 48 hr with DOX. mRNA levels were normalized by Gapdh levels and then expressed as the ratios with respect to CTR keratinocytes (shown as fold change). (D) miR-21 levels normalized to U6 in CTR (n = 4) and TG (n = 4) back skin; CTR (n = 4) and TG (n = 3) papillomas; and TG (n = 3) SCCs. (E) In situ hybrdization (ISH) of miR-21 in representative CTR and TG back skin and papillomas (bars correspond to 50 μm). (F) IHC of phospho-STAT3 (Tyr705) and phospho-AKT (Ser473) in representative CTR and TG papillomas (bars correspond to 50 μm). (G) Immunoblots of phospho-ERK (Thr202/Tyr204) and phospho-AKT (Ser473) in lysates from back skin, papillomas and SCCs from CTR and TG mice. Total ERK and TUBULIN were used as a loading controls. Bars in (B,C,D) correspond to mean ± SD of the indicated number of samples (n), each from different mouse. Statistical significance was determined by the two-tailed Student’s t test: n.s., non significant; (*) P < 0.05; (**) P < 0.01; (***) P < 0.001. In (B), statistic analysis is performed relative to CTR papillomas.
