Evaluating Invasiveness in Cancer Cells

WT and ANXA1 KO MIA PaCa-2 and HUVEC invasiveness was studied using the trans-well cell culture (12 mm diameter, 8.0-fim pore size) purchased form Corning Incorporated (New York, NJ, USA), as previously described [50 (link)]. In the lower chamber of each well were added SS, EDS, and EVs fractions from WT or ANXA1 KO MIA PaCa-2 cells as inverted media or not as technical controls. At 24 h after seeding and treatments, included the mitomycin C (10 μg/mL, Sigma-Aldrich; Saint Louis, MO, USA) to ensure the block of mitosis. The procedures of staining and analysis were performed as reported in [12 (link)].