Integrin β1 d2e5

The E-cadherin, α-tubulin, integrin α3, integrin α5, integrin β1, and β-actin amounts were determined by western blotting according to the method described in western blot analysis of ABCB1 and ABCG2 section with a slight modification. The positive control of E-cadherin, α-tubulin levels was bacitracin (1.25, 2.5, and 5.0 mM in growth medium) and it was compared to the control (growth medium). The obtained proteins were separated on 10% SDS-PAGE and visualized using a Pierce ECL Western Blotting Substrate (Thermo Fischer Scientific, USA). Proteins were detected by incubation with primary antibodies: E-cadherin (4A2) mouse mAb, α-tubulin rabbit Ab, integrin α5 rabbit Ab, integrin β1 (D2E5), β-actin rabbit Ab (Cell Signaling Technology, USA), and anti-integrin α3 rabbit Ab (St. John's Laboratory, USA) at 1:1,000 dilution in blocking buffer overnight at 4°C. β-actin was used as an internal control protein for loading normalization of the quantification analysis. The used secondary peroxidase antibodies were anti-rabbit IgG and anti-mouse IgG (Sigma Aldrich, USA). Proteins were expressed as the percentage of the controls.