Figure 1.

Effect of acriflavine (ACF) on epithelial-to-mesenchymal (EMT) induction in Panc-1 cells and invasion in vitro. (A–C) Morphological appearance of Panc-1 cells: (A) untreated, (B) after treatment with TGF-β1 (5 ng/mL) or (C) when treated with the combination of TGF-β1 (5 ng/mL) and ACF (2.5 µM). Treatment for 48 hours, original magnification is 20×, adapted from Dekervel J, Bulle A, Windmolders P, et al. Acriflavine inhibits acquired drug resistance by blocking the epithelial-to-mesenchymal transition and the unfolded protein response. Transl Oncol. 2017;10(1):59–69., Copyright (2017), with permission from Elsevier.²⁰ (D) Two-way hierarchical clustering of gene expression of Panc-1 cells treated with ACF or TGF-β1 (=T) or a combination (T-ACF) (GSE82299). A panel of 55 EMT-related genes was used (from IPA, Supplementary Table 1). The color scale illustrates the relative gene expression level: red, above the mean Z-score, green below the mean. (E) To quantify epithelial vs mesenchymal phenotype of the cells we used a representative 8-gene relative EMT score (EMT-score = ∑ SNAI1+SNAI2+TGFB1+VIM+ZEB2-CDH1-CTNNB1-AKT2, using normalized, 2log expression values, see also Supplementary File). (F) Invasion assay studies. Cells were added on top of a matrigel insert. After 48 hours the number of invaded cells was scored under a microscope on five random fields per insert (± SD).

Abbreviations: CONT, untreated control; ns, not significant.