Whole-cell protein extracts were prepared as formerly described [37 (link)]. Primary antibodies against ENG (1:1000, #14606, Cell Signal, Danvers, MA, USA) and GAPDH (1:1000, Santa Cruz Biotechnology, Dallas, TX, USA) were combined with appropriate peroxidase-coupled secondary antibodies (Jackson ImmunoResearch, West Grove, PA, USA) and employed with an enhanced chemoluminescence system (Perkin Elmer, Waltham, MA, USA).
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