Briefly, nasopharyngeal carcinoma CNZ-2Z and HNE-1 cells were collected upon oridonin treatment, and the cell lysates were extracted with protein lysis buffter. After centrifugation and denaturation, the extracts containing about 30-60 µg were subjected to SDS-PAGE (10 % or 15 % gels) and transferred onto polyvinylidene fluoride membranes (Millipore, Bedford, MA, USA). Membranes were blocked with 5% non-fat milk for 1 h and then probed with primary antibodies against E-cadherin, Vimentin, Twist1, AKT, p-AKT, STAT3, p-STAT3 and β-actin overnight at 4 °C. Subsequently, the membranes were washed with TBST buffer and incubated with horseradish peroxidase (HRP)-conjugated IgG antibody for 1 h at room temperature. Finally, the blots were detected using ECL Substrate and exposed to X-ray film according to the manufacturer's protocol.

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