Anti phf14

Western blot analyses were performed according to standard protocols as previously described [11 (link)].
Anti-E-cadherin, Anti-N-cadherin, Anti-vimentin and Anti-PHF14 were purchased from Sigma.

Proteins were extracted in radioimmune precipitation assay (RIPA) buffer (Cell Signaling, Danvers, MA, USA) containing a protease inhibitor cocktail (Roche Molecular Biochemicals, Indianapolis, IN, USA). Protein concentrations were determined by DC protein assay reagent (Bio-Rad, Hercules, CA, USA) and extracts resolved by SDS/PAGE, and transferred to PVDF membranes (Bio-Rad, Hercules, CA, USA). Membranes were then blocked for 2 h at room temperature in 0.1% TBS-Tween-20 containing 5% nonfat dried milk, washed, and then incubated with the specific primary antibodies, anti-PDGFRα (#3164, Cell Signaling Technology, Danvers, MA, USA); Anti-Actin (sc-1616, Santa Cruz Biotechnology Inc., Dallas, TX, USA); anti-PHF14 (HPA000538, Sigma-Aldrich). After washing, membranes were incubated with HRP-conjugated secondary antibodies (Santa Cruz Biotechnology Inc., Dallas, TX, USA) and proteins visualized using a Super Signal Chemiluminescence Assay kit (Pierce, Grand Island, NY, USA). The results shown were representative of three independent experiments.