FIGURE 7: AuNPs reverse EMT induced by gemcitabine on pancreatic cancer cells. Pancreatic cancer cell lines PANC-1, AsPC-1 and HPAF-II were first serum-starved for 24 h and were then divided into four groups for the treatment under the serum-starved condition: 1) NT group, 2) treated with gemcitabine (100 nM) for 72 h, 3) treated with AuNPs (25 μg/ml) for 24 h, 4) first treated with AuNPs (24 h) and then with gemcitabine (100 nM, 72h). (a) Western blot analyses of EMT markers following the afore mentioned treatment. GAPDH (1:10000) was used as the loading control. E-cad, E-cadherin (1:1000); N-cad, N-Cadherin (1:1000); Vim, Vimentin (1:1000). (b) AuNPs reverses the gemcitabine-induced relocalization of E-Cadherin in HPAF-II. At the end of the treatment, cells were fixed, stained with anti-E-cadherin antibody (1:200) followed by Alexa Fluor 488-conjugated secondary antibody (1:500), then stained with the Alexa Fluor 568-Phalloidin (1:1000) and DAPI. The localization of E-Cadherin, F-actin and nuclei were visualized by immunofluorescence microscopy. Independent experiments were repeated for at least three times.