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.

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.