Total RNA was isolated using the RNeasy Mini Kit (Qiagen) from human CD4+ CD45R0 T cells cultured in Iscove’s modified Dulbecco’s medium with KnockOut Serum Replacement (both from Life Technologies) for 17 hours in the presence of IL-1β, IL-6, IL-23, and TGF-β1 (all at 10 ng/ml) and Human T Cell Activator CD3/CD28 Dynabeads (Life Technologies). Complementary DNA was transcribed using the Reverse Transcription System (Promega). For quantitative PCR, reactions were performed using TaqMan primers and probes (Applied Biosystems). Probes used are listed in table S3. The comparative threshold cycle (CT) method (ΔΔCT) was used for relative quantification of gene expression.

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.