BT549, CAL-120, and MDA-MB-468 cells were preincubated in DMEM-F12 or Plasmax (both supplemented with 2.5% FBS) for 2 days for the experiments reported in Fig. 1, and for all experiments, cells were seeded in six-well plates in 2 ml of medium. Cell densities, incubation times, and components added to the media are indicated in Figs. 1 and 2. Fresh medium was added after 96 hours to prevent nutrient exhaustion. At endpoint, colonies were fixed with trichloroacetic acid (final concentration of 3%, 30 min incubation at room temperature), washed with water, and stained with sulforhodamine B (0.057% in 1% acetic acid, 1 hour incubation at room temperature). LI-COR Odyssey and Image Studio 5 were used for image acquisition, and ImageJ was used for quantification of area covered by colonies and colony number. For the identification of sodium selenite as a stimulator of colony formation, MDA-MB-468 cells were cultured in a mixture of DMEM-F12 and Plasmax (1:1) and in DMEM-F12 supplemented with the different Plasmax stock solutions or individual Plasmax components reported in table S1.

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



Q&A
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.