Figure 3
Differential expression of EMT- and stemness-related genes and proteins in MI6 and WTHER2_1 cells and invasiveness properties. (a) Analysis of EMT and stem cell-related genes differentially expressed (fold) in MI6 vs WTHER2_1 cells as shown by the Mouse EMT RT22 Profiler PCR Array. Changes in gene expression were analyzed by SABioscience software (SABiosciences, Qiagen, Hilden, Germany) using GAPDH for normalization. (b) Quantitative reverse transcriptase (qRT)–PCR validation of the differential expression of genes involved in EMT and stemness. The data are the mean±s.d. (n=2) and given as fold increase of relative expression in MI6 vs WTHER2_1 cells (MI6/WTHER2_1). (c) Western blot analyses of MI6 and WTHER2_1 protein extracts separated by 4–12% gradient SDS–PAGE under reducing conditions to evaluate SOX10, basal and activated FAK (pFAK), TGFβ, basal and cleaved NOTCH4 and WNT5A proteins. Vinculin and Actin were used to normalize protein loading. Autoradiographs were acquired at different exposure times to obtain optimal image resolution. (d) Migration and invasion capability of MI6 and WTHER2_1 cells. The results are the mean±s.d. (n=3). Significance was calculated by a two-tailed unpaired t-test.
