*2.1. Animals*

Female CD1 mice (*Mus musculus*) from the Instituto de Higiene e Medicina Tropical animal house were used to obtain blood for membrane feeding assays (MFAs) and mosquito infections, with the corresponding license (009511 from 21 April 2019) approved by the Portuguese National Authority Health (DGAV). For ex vivo production of ookinetes, female BALB/c mice (Janvier Labs, Le Genest-Saint-Isle, France) were used, following the protocols reviewed and approved by the Ethical Committee on Clinical Research from the *Hospital Clínic de Barcelona* (Reg. 10100/P2, approved on January 2018). In all cases, for the experimental procedure, mice were anesthetized using 100 mg/kg ketamine (Ketolar) mixed with 10 mg/kg xylazine (Rompun) intraperitoneally (i.p.) administered, and regularly monitored. *Anopheles stephensi* mosquitoes were maintained under standard insectary conditions (26 ± 1 ◦C, 75% humidity and a 12/12 h light/dark cycle). Adult mosquitoes were fed on 10% glucose solution ad libitum until the day before feeding trials.
