Western Blotting Procedure for Protein Analysis

Western blotting was performed using standard procedures [24 (link)]. The primary antibodies used were anti-NEDD9 (2G9), -phospho-ERK1/2(D13.14.4E), -ERK1/2(137F5), -phospho-Aurora Kinase A(D13A11), -HER2 (29D8), -phospho-Src (100F9), FAK (D2R2E) (Cell Signaling Technology, Danvers, MA USA), anti-phospho-FAK (Tyr397) (ThermoFisher Scientific, Waltham, MA, USA), anti-Src (GD11) (Sigma-Aldrich, St. Louis, MO, USA), anti-Aurora Kinase A (BD Biosciences, Franklin Lakes, NJ, USA). The dilution was based on the manufacturer’s recommendation. Secondary anti-mouse and anti-rabbit horseradish peroxidase (HRP)-conjugated antibodies (Jackson ImmunoResearch Laboratories, Inc., West Grove, PA, USA) were diluted, 1:10,000, followed by chemiluminescence-based detection with Luminata Forte Western HRP substrate (Sigma-Aldrich, St. Louis, MO, USA), and were quantified using ImageJ software (ImageJ—National Institute of Health, https://imagej.nih.gov, accessed on 12 December 2022).