Figure 4. Low Cav1 expressing cells undergo an EMT.

A. Phase contrast images of stable SCC9 cells expressing shRNA_ctrl or shRNA_cav1 in culture. B. Quantitative determination of transcripts expression of epithelial (E-cadherin and β-catenin) and mesenchymal (vimentin) markers in shRNA_ctrl- or shRNA_cav1-SCC9 cells using qRT-PCR with RNA18S as control and western blot using tubulin as a loading control. Each bar represents the mean±SEM with **p < 0.01 and *** < p0.001. C. Quantitative determination of transcripts expression of key regulator of E-cadherin and EMT (including Slug, Snail, Twist, ZEB1 and ZEB2) in shRNA_ctrl- or shRNA_cav1-SCC9 cells using qRT-PCR with RNA18S as control. Each bar represents the mean±SEM with **p < 0.01 and ***p < 0.001. D. Quantified analysis of E-cadherin, vimentin, Twist and Cav1 transcripts determined in primary tumor samples of patients that developed metastasis (“R1”) and primary tumor samples of patients that did not developed metastasis (“non-R1”). The line within the bar represents the mean value and “o” represent individual data point. (***p < 0.001). E. Quantitative determination of transcripts expression of Slug, Snail, Twist, Zeb1, vimentin and E-cadherin in shRNA_ctrl- or shRNA_cav1-SCC9 cells transfected by siRNA_ctrl or siRNA_SLUG or siRNA_TWIST using qRT-PCR with RNA18S as control. Each bar represents the mean±SEM with *p < 0.05, **p < 0.01 and *** < p0.001. F. Quantitative determination of transcripts expression of Slug, Snail, Twist, Zeb1, vimentin and E-cadherin in shRNA_ctrl- or shRNA_cav1-SCC9 cells exposed to DMSO or 20 μmol/L K34c using qRT-PCR with RNA18S as control. Each bar represents the mean±SEM with *p < 0.05, **p < 0.01 and *** < p0.001.