Western Blot Analysis of HGSOC Cell Lines

Western immunoblot analysis was performed using whole-cell lysates derived from HGSOC cell lines as previously described with minor alterations.^{15 (link)} To lyse cells grown as spheroids, 1% sodium dodecyl sulfate was added to the lysis buffer. Approximately 40 μg of protein lysate per cell line was analyzed. Equal loading was verified by incubating the membranes with anti-GAPDH antibody. Primary antibodies used include monoclonal mouse anti-GAPDH (GeneTex, Irvine, CA, USA; cat. no. GTX627408), monoclonal rabbit anti-E-cadherin (Cell Signaling, Danvers, MA, USA; cat. no. 3195), polyclonal rabbit anti-SUSD2 (Prestige Antibodies Sigma-Aldrich Corp.; cat. no. HPA004117) and polyclonal rabbit anti-N-cadherin (Cell Signaling; cat. no. 4061). Secondary antibodies used were Pierce (Thermo-Fisher Scientific, Waltham, MA, USA; cat. no. 31329) for GAPDH. Pierce cat. no. 31345 was used for N-cadherin, E-cadherin and SUSD2. Experiments were performed using three biological replicates for each cell line.