Fig. 2.

Analysis of mutations in a D. melanogaster model. (A) Design of transgene constructs. T2A is a viral sequence causing translation of two proteins. Red dots indicate amino acid changes. (M99) and (M127) label start codons after NTΔ. 1) positive CI control - the cid^wMel backbone. 2) cid^wMel (V-L) point mutant. 3) cid^wMel (NTΔ) starting at M99. 4) cid^wYak analog, in effect, wYak wild-type. 5) positive CI control, the cin^wPip backbone. 6) cin^wYak analog, in effect, wYak wild-type. 7) pUASp-attb insertion vector. (B) Transgenic CI with the weak NGT driver. Males expressing cid^wMel cause strong CI relative to uninfected (U). Males expressing cid^wMel(V-L) cause hypomorphic CI. wMel cytoplasmic infection rescues CI from males expressing cid^wMel or cid^wMel(V-L). (C) CI analysis with boosted expression using an MTD driver. Males expressing cid^wMel or cid^wMel(V-L) cause strong CI that cytoplasmic wMel infections in females fully rescue. Males expressing cin^wPip cause weak CI. (D) Transgenic rescue. wMel cytoplasmic infection in males causes strong CI. wMel cytoplasmic infection in females rescues wMel-induced CI. Expression of operons cid^wMel or cid^wYak in females fully rescues wMel-induced CI. Raw hatch rates are plotted over means and SDs. (*) is P < 10⁻⁶, and (ns) is not significant. P values for B and C are calculated from one-tailed Wilcoxon tests with Bonferroni correction for multiple comparisons, while P values for D are calculated by a Kruskal–Wallis analysis with Dunn’s multiple comparison test.