To evaluate the cellular uptake of Dox conjugates, MMTV-PyMT cells were seeded in a 96-well high-content imaging plate (Corning) at 5000 cells per well and incubated overnight. Cells were washed with PBS and treated with free Dox, Dox-SA, or Dox-CBD-SA dissolved in DMEM [sodium pyruvate (110 mg/liter), 10% heat-inactivated FBS, 1% penicillin-streptomycin, and Phenol red (−)] at a concentration of 50 μM equivalent of Dox. After incubation, cells were washed twice, treated with 75 nM LysoTracker Deep Red, and further incubated for 30 min at 37°C. Cells were washed twice and observed by an IX83 microscope (Olympus) with ×60 magnification. Images were processed using ImageJ software (National Institutes of Health, NIH). Scale bar, 20 μm.

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.