X single cell analysis software

Synchronous ring-stage or late-stage infected red blood cells (iRBC) and uninfected red blood cells (uRBC) were brought to 4% hematocrit in complete culture medium. Fluorescein-conjugated peptides were added to a final concentration of 1 μM, and cultures were incubated for 6 h at 37 °C under standard gas conditions. Following incubation, 25 μL of cell mixture was removed and stained with 100 μL of 2 μg/mL Hoechst 33342 for 10 min at 37 °C. Cells were subsequently washed once in 1 mL of 1× PBS, resuspended in 300 μL of 1× PBS, and analyzed for Hoechst and fluorescein staining on a Beckman Coulter CyAn flow cytometer. 500 000 events were collected at a rate of 15 000–20 000 events per second. Data was analyzed using FlowJo X single cell analysis software (FlowJo LLC).