Cells were plated at equal concentrations with varying concentrations of the inhibitor and in triplicate wells. Cells were counted, the concentration was calculated every second/third day, and the cells were replated at even cell concentrations. A growth curve was plotted for cumulative growth according to the dilution at each time point. Doubling time was calculated from fitted semi-logarithmic slope (Nonlin). Fit using least squares in GraphPad Prism software by log2/slope and Student’s two-tailed t test was used to test for significance. For cell cycle assays (Lp30 cells), cells were plated with 30 μM inhibitor or DMSO for 24 hours. Cells were harvested, and an equal number of cells were fixed in 2% paraformaldehyde, permeabilized in 0.1% saponin, stained with 4′,6-diamidino-2-phenylindole (DAPI), and analyzed by flow cytometry (LSR-II, BD Biosciences). For apoptosis assay (Lp30 cells), cells were plated with 30 μM inhibitor or DMSO for 72 hours, harvested, washed and stained using a PE-annexin V kit (BD Biosciences), and analyzed by flow cytometry (Accuri, BD Biosciences). All flow cytometry analyses were run with FlowJo software V.10.

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.