Dmbl microscope

Drug sensitivity assays against mature (stage V) P. falciparum gametocytes were performed as described previously (Lelièvre et al., 2012 (link); de Carvalho et al., 2019 (link)). Briefly, the drugs were precoated in a 3-fold dilution in 96-well plates. The concentration of the solvent did not interfere with parasite growth. Epoxomicin and methylene blue (Supplementary Table 1) were used as positive control. Subsequently, the previously purified mature gametocytes were seeded to the plates (50,000 gametocytes/well) and incubated at 37°C in 5% CO₂ and 5% O₂. After 48 h, the ATP production was measured by the BacTiterGlo assay (Promega), according to the manufacturer’s instructions. Finally, the results were quantified using a luminometer (LUmo; Autobio). In addition, to visually inspect the effect of boromycin on gametocytes thin blood smears of untreated and boromycin (8 nM) and epoxomicin (15 nM) - treated gametocytes (50,000 gametocytes/well), were prepared, fixed in 100% methanol for 10 seconds, and stained with Giemsa (Merck) solution (5%) for 20 minutes. The slides were washed and observed under a Leica DMBL microscope, and pictures were taken using a ProgRes C10 camera and software (Jenoptik), at 100X magnification.