*2.3. Membrane Blood Feeding Assay*

Blood obtained from an intracardiac puncture of a CD1 mouse infected with *P. berghei* ANKA-GFP (259cl1; MRA-865) was treated with 1/10 volume of 3.2% *w*/*v* sodium citrate to prevent coagulation. 6.25 µL of a solution prepared by dissolving heparin at 40 mg/mL or 0.4 mg/mL in phosphate buffered saline (PBS) was added to 494 µL of blood:citrate (to obtain final heparin concentrations of 500 and 5 µg/mL, respectively), which was then placed in feeders prepared with two-sided stretched Parafilm® connected to two plastic tubes for water inlet and outlet. The same volumes of PBS and blood:citrate were used for heparin-free controls. Temperature within the multiple cylindrical water-jacked glass was kept at 37 ◦C by a constant water flow supply. Each feeder was placed on top of a net-covered paper cup containing 40–50 *A. stephensi* females. Mosquitoes were allowed to feed for one hour. Non-fed mosquitoes were removed, and the rest were treated as above. The final number of mosquitoes analyzed for the non-modified heparin assay was 127 in the control group, 106 treated with 5 µg/mL heparin, and 149 treated with 500 µg/mL heparin, distributed in three independent experiments. The final number of mosquitoes analyzed for the hypersulfated heparin assay was 62 in the control group, 91 treated with 5 µg/mL hypersulfated heparin, and 102 treated with 500 µg/mL hypersulfated heparin, distributed in two independent experiments.
