Fig. 6. ER stress upregulates EMT markers in PC CL3 cells. (A) RT-PCR analysis of total RNA extracted from PC Cl3 cells vehicle treated or treated with 0.5 µM TH or 0.5 µg/ml TN for 30 minutes, followed by 24 and 48 hours in medium without TH or TN. (B) Western blots of total protein extracts from PC Cl3 cells vehicle treated or treated with 0.5 µM TH or 0.5 µg/ml TN for 30 minutes, followed by 24 and 48 hours in medium without TH or TN. Filters were probed with antibodies against E-cadherin and β-actin (control). (C) Real-time RT-PCR analysis of total RNA isolated from PC Cl3 cells vehicle treated or treated with 0.5 µM TH or 0.5 µg/ml TN for 30 minutes, followed by 24 hours in medium without TH or TN. Each bar represents the mean±s.d. of four independent experiments, each performed in triplicate. β-actin was used as an internal standard (β-actin values were not affected by TH or TN treatments) (REU, relative expression units). Asterisks indicate statistically significant differences (^***, P<0.001). (D) RT-PCR analysis of total RNA extracted from PC Cl3 cells vehicle treated or treated with 0.5 µM TH and 0.5 µg/ml TN for 30 minutes, followed by 6, 24 and 48 hours in medium without TH or TN.