Figure 3

Upregulation of Sox2 in SIX1-overexpressing tumors. (a) Volcano plot of RNASeq results. FC, expression fold change in SIX1-tumors relative to controls; -log padj, minus logarithm of the adjusted p value. The horizontal dotted line indicates padj = 0.05, the vertical dashed lines indicate FC 2 and −2. (b) QPCR analysis of Sox2 expression in tumors with (SIX1) or without (V) SIX1 overexpression, (V/RAS n = 4, SIX1/RAS n = 6). (c) Immunohistochemical detection of Sox2 in the indicated tumors. The left panel shows a representative staining and the right panel shows a quantification of positive cells (n = 6 for each tumor type). Main scale bar, 50 µm; inset scale bar, 20 µm. (d) Immunofluorescence of SIX1, Sox2 and p16Ink4a in the indicated tumors. Scale bar, 100 µm. (e) Flow cytometry detection of the activity of the Sox2-responsive SORE6 cassette in the indicated fibroblasts transduced with SORE6 or empty vector (CMV).