Figure 5

LD enrichment involves fatty acid synthesis and storage in the form of triacylglycerol.

Aag2 cells were incubated or not with heat killed Ec and 24 hours later lipids were extracted and either fractioned using a TLC for neutral lipids (A) or cells were lysed and subjected to enzymatic determination of triacylglycerol contents (B). Alternatively, Aag2 cells were incubated with heat killed Ec in the presence or absence of 10 μg/ml of C75 (a Fatty acid synthase inhibitor). After 2  hours cells were lysed and subjected to enzymatic determination of triacylglycerol contents (C). In a different experiment, Aag2 stimulated or not with heat killed Ec (D–F) or midguts from mosquitoes fed only on sugar or with blood (G–I) had their total RNA extracted and the expression of Fatty acid synthase (FAS) (D,G), Lipid storage droplet −1 (LSD-1) (E,H) or Lipid storage droplet −2 (LSD-2) (F,I) genes was determined by qPCR.