iSLK/RTA-WT and iSLK/RTA-DD stable cell lines were established using Tet-On 3G Tetracycline Inducible Gene Expression Systems. Briefly, 293T cells were transfected with the packaging plasmids VSV-G, Gag-pro, Vpr-pol, Tet-off, and tat-IRES-rev and the TetONE-RTA-WT or TetONE-RTA-DD lentiviral expression vector. For making shRNA lentivirus, 293T cells were cotransfected with packaging plasmids VSV-G, DR8.9, and the shRNA lentiviral plasmid as previously described (53). At 48 hours after transfection, the supernatant was harvested and filtered, and viruses were concentrated by ultracentrifugation. SLK or 293T cells were spin-infected (1800 rpm) with lentivirus in the presence of polybrene (8 μg/ml) for 45 min. Cells were selected with puromycin (1 μg/ml) at 48 hours after infection.

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.