Improve Research Reproducibility A Bio-protocol resource
- Home
- Protocols
-
Request a Protocol
Ask a
question
Favorite
For transient expression, cells were transfected with lipofectamine 2000 (Invitrogen) or Mirus X2 (Mirus Bio. LLC) at approximatively 60% confluency. For infections, Phoenix cells were transfected at approximatively 50% confluency using lipofectamine 2000, with 10 μg of retroviral plasmids (pMSCV-puromycin) and 5 μg of packaging plasmid. Virus-containing supernatant were collected, filtered (0.45 μm), and added to target cells (HCT116, hTERT-RPE1) with polybrene (8 μg/ml). Infected cells were selected on puromycin (HCT116, 2 μg/ml; hTERT-RPE1, 5 μg/ml).
Copyright and License information: The Author(s) ©2021
Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
Do you have any questions about this protocol?
Post your question to gather feedback from the community. We will also invite the authors of this article to respond.
Post a Question0 Q&A
