Ix71 microscopy system

Uniform wounds were introduced into cell cultures using a culture insert (#80201; ibidi, Munich, Germany). The inserts were placed in individual wells of a six-well plate. Ishikawa, HEC-1B and AN3CA cells transfected with vector for 96 h or AN3CA cells treated with sitagliptin or left untreated were seeded in each reservoir of the insert (1×10⁴cells in a finalvolume of 100 μl). Inserts were removed after cells adhered. The gaps (wounds) were washed with serum-free medium, and 2ml fresh medium were added to each well. AN3CA cells were treated with 1 mMsitagliptin, with culture medium aloneserving as a control. Cell migration into the wound area was recorded using an IX71 microscopy system coupled to a DP73 digital camera (both from Olympus, Tokyo, Japan) at 0 and 48 h. Wound closure at 24–48 h was compared with time 0.

We confirmed ER1 and PR1 expression in all EC cell lines via qRT-PCR as compared with analogous data provided by ATCC. EC cells morphologies before and after DPPIV up- or downregulation were also recorded using an IX71 microscopy system coupled to a DP73 digital camera (both from Olympus, Tokyo, Japan).