Figure 4.

Role of Two Midgut Peroxidases on Prostaglandin Synthesis and Immune priming

(A) Relative mRNA expression of HPX7 and HPX8 in the midgut, 48 h post blood-meal, in LacZ (control) and IMPer-silenced mosquitoes.

(B) Effect of IMPer and HPX7 co-silencing on hemolymph prostaglandin levels 48 h after blood feeding.

(C) Effect of IMPer and HPX8 co-silencing on hemolymph prostaglandin levels 48 h after blood feeding.

(D and E) Effect of IMPer, HPX8, or IMPer + HPX8 co-silencing on the number of hemocytes attached to the basal surface of the midgut 12 h post blood feeding. Actin (phalloidin), cyan; Hemocytes (stained with Vybrant CM-DiI), red. Scale bar: 15 μm.

(F) Effect of hemolymph transfer from LacZ controls and HPX7-silenced donors, collected 4 days after feeding on a control (C) or a P.berghei infected (I) mouse, on the proportion of circulating hemocytes in the recipients.

(G) Effect of hemolymph transfer from LacZ controls and HPX8-silenced donors, collected 4 days after feeding on a control (C) or a P.berghei-infected (I) mouse, on the proportion of circulating hemocytes in the recipients.

(H) Effect of hemolymph transfer from LacZ controls and HPX8-silenced donors on the antiplasmodial immunity of the recipients. Hemolymph was collected 4 days post-feeding on control (C) or a P.berghei-infected (I) mouse.

Error bars in A–C, F, and G represent mean ± SEM. Mann-Whitney U test and Unpaired t test, **p ≤ 0.01; ***p ≤ 0.001, NS, p > 0.05. In A–C, 2–3 pools per treatment were used in at least two independent experiments. In F and G, granulocyte numbers were counted in at least 5–10 individual mosquitoes. Each dot in E and H represent the number of hemocytes or oocysts, respectively, for each individual midgut. The median is indicated by the line. Detailed information on biological replicates and exact p values are listed in Tables S20–S26.