Figure 2

Synergistic effect of vactosertib with nal-IRI/5-FU on migration, invasion, and EMT of pancreatic cancer cells. (A) Transwell cell migration assay. Cells were treated with the indicated reagent(s) and incubated for 48 hours before placed in a migration chamber. 1 × 105 of PANC-1 (left) or 5 × 104 of Panc02 (right) cells were then seeded in migration chambers for 48 hours. The relative number of migrated cells in each group was counted. (B) Cell invasion assay. Cells were treated with the indicated reagent(s) and incubated for 48 hours before placed in an invasion chamber. 2 × 105 PANC-1 (left) or 1 × 105 Panc02 (right) cells were then seeded in invasion chambers for 48 hours. The number of invaded cells in each group was counted. Note that the combination of vactosertib with nal-IRI/5-FU inhibits migration and invasion in PANC-1 and Panc02 pancreatic cancer cells. (A,B) The values for migrated/invaded number of cells represent the mean ± SD of triplicate data. ***P < 0.0005, **P < 0.005, and *P < 0.05 compared to the control group; ##P < 0.005 and #P < 0.05 compared to the nal-IRI-treated group. (C) Western blot analysis for EMT markers. Vactosertib was treated for 2 hours prior to incubation with nal-IRI/5-FU in the presence of 5 ng/ml TGF-β for 48 hours. The blots are cropped and the full-length images are presented in Supplementary Fig. 8. Note that vactosertib and its combination with nal-IRI/5-FU restore the TGF-β-mediated reduction of E-cadherin and the increase of mesenchymal markers.