A culture containing uRBCs and 5–10% late-stage (trophozoite/schizont) iRBCs was washed twice in warm Ringer Solution (125 mM NaCl, 5 mM KCl, 1 mM MgSO4, 32 mM HEPES, 5 mM glucose, pH 7.4). RBCs were resuspended in Ringer Solution with 2.5 mM CaCl2, and incubated for 5 minutes with 0.5 mM sodium orthovanadate, 2 μM A23187 (Calbiochem), or the corresponding volume of buffer only. 5% v/v FITC-Annexin V stain was added to triplicate wells of each treatment, and incubated for 20 minutes at 37°C. Samples were resuspended in Ringer Solution (2.5mM CaCl2) with 5 μg/mL Hoechst 33342 and incubated for 20 minutes at 37°C. Images and flow cytometry data were collected and processed as described above. The percentage of uRBCs (Hoechst-negative) and iRBCs (Hoechst-positive) populations with FITC-Annexin V binding was calculated in FlowJo by setting a cut-off above unstained controls (S3A Fig). Data were analysed as specified.

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.