Most experiments were carried out with Aedes aegypti mosquitoes derived from a wild population originally sampled in 2013 in Thep Na Korn, Thailand and took place within 10 generations of laboratory colonization. One experiment was carried out with Ae. aegypti mosquitoes derived from a wild population originally sampled in 2015 in Phnom Penh City, Cambodia and took place 8 generations after laboratory colonization. Experimental infections were carried out as previously described [60 (link)]. Briefly, four- to seven-day-old females were offered a washed rabbit erythrocyte suspension mixed 2:1 with pre-diluted DENV-1 KDH0030A viral stock and supplemented with 10 mM ATP (Sigma), to reach an expected titer of 107 FFU/mL. A control blood meal was prepared with the supernatant of mock-inoculated C6/36 cells. Mosquitoes were allowed to blood feed for 30 min through a pig-intestine membrane using an artificial feeder (Hemotek Ltd, Blackburn, UK) set at 37°C. Samples of the blood meals were saved and stored at -80°C for further titration. Fully engorged females were incubated at 28°C, 70% relative humidity and under a 12-hour light-dark cycle in 1-pint cardboard cups (20–30 females per cup, at least 2 cups/condition) with permanent access to 10% sucrose.
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