8 well tissue culture chamber slides

Ovarian cancer cells were grown on an 8-well tissue culture chamber slides (Sarstedt, Switzerland), fixed with 4% paraformaldehyde, permeabilized with 0.3% Triton X100, and blocked with 5% (w/v) FBS (Sigma-Aldrich), 1% bovine serum albumin (BSA) fraction V, and 0.1% TritonX-100 containing PBS for 1 h at room temperature. Cells were then stained with anti-integrin α2, anti-E-cadherin antibodies for overnight at 4°C. Following extensive washing, corresponding secondary antibodies were added to each chamber and incubated for 2 h. Cells were washed with PBS containing 0.1% Tween 20 and incubated and counterstained with ProLong® Gold antifade reagent with DAPI (Cell Signaling Technology). Fluorescence images were taken by a Zeiss LSM 710 confocal microscope (Zeiss, Feldbach, Switzerland). For patient-derived primary cells, 100-200 μL ascites cell suspension was collected using cytospin for 5 min at 400g on a glass slide. Cells were then fixed with 4% paraformaldehyde, permeabilized, blocked and stained as mentioned above.

Ovarian cancer cells were grown on an 8-well tissue culture chamber slides (Sarstedt, Switzerland), fixed with 4% paraformaldehyde, permeabilized with 0.3% Triton X100, and blocked with 5% (w/v) FBS (Sigma-Aldrich), 1% bovine serum albumin (BSA) fraction V, and 0.1% TritonX-100 containing PBS for 1 hr at room temperature. Cells were then stained with anti-integrin α2, anti-integrin β1, or anti-E-cadherin antibodies for overnight at 4°C. Following extensive washing, corresponding secondary antibodies were added to each chamber and incubated for 2 hr. Cells were washed with PBS containing 0.1% Tween 20 and incubated and counterstained with ProLong Gold antifade reagent with DAPI (Cell Signaling Technology). Fluorescence images were taken by a Zeiss LSM 710 confocal microscope (Zeiss, Feldbach, Switzerland). For patient-derived primary cells, 100–200 μL ascites cell suspension was collected using cytospin for 5 min at 400 g on a glass slide. Cells were then fixed with 4% paraformaldehyde, permeabilized, blocked, and stained as mentioned above.