Fig. 3
Mosquito lipids regulate within-vector parasite sporogony. Control and Lp-deficient female mosquitoes were infected with P. berghei (P.b.) (a–c) or P. falciparum (P.f.) (d, e) parasites. a Neutral lipids detected by the Nile Red stain in the GFP-expressing P. berghei oocysts (green) 13 day post infection (dpi) in control (Ctrl) and Lp-depleted (dsLp) mosquitoes. Nuclei are visualized by DAPI (blue). Scale bars—10 µm. b Proportion of Nile Red-positive P. berghei oocysts in control and Lp-depleted mosquitoes 13 dpi (N = 3, 21 mosquitoes per condition per experiment). Each dot represents mean of one experiment (*p < 0.05; paired one-sided t-test, paired values for dsRNA treatment per experiment). c Oocysts detected by transmission electron microscopy (Supplementary Methods) were scored according to the developmental stage: non-sporulating oocysts with normal morphology (immature, white); oocysts with normal plasma membrane retraction, budding sporoblasts, or formed sporozoites (sporulating, gray); aberrant oocysts with electron-light intracellular vacuolization (abnormal/dead, black). Proportions of the categories are shown as pie charts. The total number of analyzed oocysts per condition is indicated in the middle of each chart (N = 1 (7 dpi) and N = 3 (13 dpi), 5–14 mosquitoes per condition per experiment). The results of one representative experiment are shown. d Oocyst sizes of P.f. (11 dpi, N = 3) and P.b. (14 dpi, N = 6) in control and Lp-depleted mosquitoes (5–31 mosquitoes per condition per experiment). Measurements of individual mosquitoes and experiments were pooled (total numbers of oocysts are shown in brackets). The boxplot shows the median (center line), the 25th to 75th percentiles (bounds of box) and the 10th and 90th percentiles (whiskers) (***p < 0.0001; one-sided Mann–Whitney test). e Development of P.f. (14 dpi, N = 6) and P.b. (18 dpi, N = 6) salivary gland sporozoites in control and Lp-depleted mosquitoes (3–75 mosquitoes per condition per experiment). Each dot represents the mean number of sporozoites per mosquito per experiment. (P.f.) *p < 0.05, paired one-sided Wilcoxon matched-pairs signed rank test; (P.b.) ***p < 0.0001 paired one-sided t-test, paired values for different dsRNA treatment per experiment
