Figure 7—figure supplement 3.

(A) IGROV1 WT and ΔITGA2 cells were treated with 0, 1, or 10 μM of Defactinib for 16 hr and stained with Annexin V-FITC and DAPI to identify apoptotic dead cells. Data represent the mean ± SD (*p<0.05) from two independent experiments. (B) Anchorage-independent soft agar assay, cells were incubated with or without 5 μM of Defactinib for 10 days. Scale bar 100 μm. (C–D) Bar chart shows the quantification of total colony and the average diameters of spheroids (***p<0.001). (E) Cancer cell-collagen adhesion assay. Representative images show SKOV3 cell adhesion to collagen in the presence of DMSO, 1 μM of TC-I-15, Defactinib (FAK inhibitor) and FR180204 (ERK1/2 inhibitor). Bar chart with mean ± SD of the percentage of cell adhesion efficiency of cancer cells. (unpaired Student’s t-test, ***p<0.001). (F) Mesothelial clearance assay. SKOV3 cancer spheroids were seeded on top of a GFP-expressing MeT-5A mesothelial monolayer in the presence of DMSO or inhibitors for 20 hr. Mesothelial clearance area was measured by the size of black hole (white dash circle) using Image J software. Bar chart represents mean ± SD of clearance area from two independent experiment. Scale bar 200 μm.