Lentiviral constructs with shRNAs were directed against murine Ms4a6d, A20, and Stat3 in the pGV112 vectors. The primers are shown in table S1. Lentivirus was prepared by transient transfection of 293T cells with transfer vectors along with third-generation packaging constructs (pHelper 1.0 and pHelper 2.0). The viral titers were determined with serial dilution of virus-containing media on NIH3T3 cells. RAW264.7 cells or PEMs were transfected with unconcentrated virus supernatant overnight in the presence of polybrene (8 mg/ml) and selected in puromycin (0.5 mg/ml).

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.