Western blot was performed as previously described [31 (link)]. Briefly, a radioimmunoprecipitation assay (Cell Signaling Technology) enhanced with cOmplete™ protease inhibitor cocktail (Roche Applied Science), 1 mM sodium fluoride, 2 μg/ml aprotinin, and 1 mM phenylmethylsulfonyl fluoride was used to lyse the prostate cells. Quantification of protein was assessed by a Bradford assay. Primary antibodies, such as Cldn3 and Cldn4 (Novus), and secondary antibodies, such as anti-mouse and anti-rabbit (Cell Signaling Technology), were used. Protein signal was detected using Chemiluminescence (Thermo Scientific, Rockford IL).
Free full text: Click here
