Figure 1

Experimental design. For each of the four lines of Anopheles gambiae – black refractory (BR), black susceptible (BS), red refractory (RR) and red susceptible (RS) – four sub-lines were established with the same genetic background (only shown for the BR line) by dividing ~400 pupae from 8 first generation (G1) females among four replicate cages (A, B, C and D). The adults in the second generation (G2) produced the males (sires) used in the assay of male reproductive success. The G2 adults from the Keele population produced the females (dams) used in the assay of male reproductive success. All third generation (G3) larvae were reared individually in the wells of 24-well tissue culture plates. In the third generation (G3), a cage of 28 males was obtained for each of the 16 sub-lines (only shown for cage A of the BR line). Each cage of males was mated at 3 and 5 days of age to two different groups of 15 virgin Keele females (aged 1 and 3 days, respectively). Males and females were allowed to mate for 20 hours for each mating day. The females were blood fed two days post-mating (pm). Ten blood fed females were haphazardly selected and allocated to individual oviposition cups 3 days pm. All females were given a second blood meal (8 days pm), were monitored for oviposition (3 – 13 days pm) and were dissected for their spermathecae (14 days pm).