<i>Wolbachia</i> density by treatment group.

<p><i>Wolbachia</i> densities in 0–2 d-old (circles) and 4–7 d-old (inverted triangles) female <i>w</i>Mel <i>A</i>. <i>aegypti</i> exposed to various temperature treatments. “<i>w</i>Mel” denotes the <i>w</i>Mel-infected <i>A</i>. <i>aegypti</i> controls that were not exposed to high temperatures, and for the other treatment groups the letter represents the stage of heat onset—“E” for embryogenesis, “I” for 1^st/2^nd instars, and “M” for 3^rd/4^th instars—and the number represents the number of days the group remained in the high temperature treatment. <i>Wolbachia</i> density was measured by qPCR of the <i>Wolbachia</i>-specific <i>wsp</i> gene and the somatic insect gene <i>Actin5c</i>. Displayed values are relative concentrations of <i>wsp</i> and <i>Actin5c</i> calculated in Q-Gene. The horizontal line at <i>y</i> = 10^−3.552 represents the detection limit of <i>Wolbachia</i> by qPCR, which was established by the average C_q values for <i>Wolbachia</i>-free Cairns <i>A</i>. <i>aegypti</i> controls. Bars denote means bounded by their 95% confidence intervals. The lower 95% confidence limit for the 0–2 d-old I5 group (y = −0.009) is not shown because it cannot be represented on the log scale. The significance levels of differences between time points are indicated above brackets and between treatment groups and the <i>w</i>Mel controls at the top of the graph as <i>P</i> < 0.05 (*), <i>P</i> < 0.01 (**), <i>P</i> < 0.001 (***). Each point represents an individual mosquito.