Figure 2.

A: Western Blot analysis of AKT pathway and EMT related proteins. Bar graphs showed the results of densitomatric analysis (mean ± standard deviation) (* indicates P<0.05, student t-test). Phosphorylated and total protein for EGFR, AKT, ERK, PI3K were increased in arsenic treated HUC1 cells. CyclinD3 and mTOR were also increased in arsenic treated HUC1 cells. The molecules that showed significant alterations due to arsenic treatment were bolded. As: arsenic; B: Expression levels of miR-200 family in arsenic exposed HUC1 cell culture model. HUC1 cell line was cultured in the presence of 1µM As₂O₃ for 6, 8 and 10 months. In order to study the reversibility of the arsenic exposal effect to the cells, we withdraw the drug for 2.5 months. Using the 2^−ΔΔCT method, the data are presented as the fold change in each miRNA expression normalized to mir-222 and relative to the untreated control (HUC1 cell line cultured in the presence of PBS for the same time course). Statistical difference between arsenic treated and untreated cells were calculated using Student’s t-test. * indicates P<0.05. C: Western blot analysis of two EMT associated protein: E-cadherin and Vimentin expression were analyzed using arsenic treated and untreated HUC1 cells for 6 months and 10 months. E-cadherin was significantly decreased in Arsenic treated cells of both 6 and 10 months (P<0.001 for both 6 and 10 months, Student’s t-test); No changes of expression was observed for Vimentin at 6 months arsenic treated cells, however, Vimentin expression dramatically increased at 10 months Arsenic treated cells (P<0.001, Student’s t-test).