Table 1 Description of three different MALDI mass spectrometry protein-extraction protocols used in the present study

Selected mosquito body parts were placed in separate microcentrifuge tubes, rinsed with 300 μL ultrapure water and 900 μL ethanol, and centrifuged at 13,000 rpm for 2 min

Samples were decanted and treated with 10 μL of 70% formic acid for 5 min at room temperature

Immediately after, samples were homogenized in the tube with the help of a manual pestle with an additional 10 μL of 100% acetonitrile and centrifuged at 13,000 rpm for 2 min

A small volume of supernatant was pre-mixed with equal volume of 10 mg/mL α-cyano-4-hydroxycinnamic acid (HCCA) matrix and 1 μL of the mix was quickly placed in its respective target well in triplicate

Selected mosquito body parts were rinsed with distilled water and dried with paper

Samples were immediately homogenized with the help of a manual pestle in 20 μL of 70% formic acid and 20 μL of 100% acetonitrile and incubated for 1 h

Samples were vortexed for 15 s, centrifuged at 13,000 rpm for 2 min and a small volume of the supernatant was pre-mixed with equal volume of 10 mg/mL HCCA before adding 1 μL of the mix it to the target well in triplicate

Selected mosquito body parts were homogenized with the help of a manual pestle in 20 μL of 10% formic acid, pre-mixing with 1.5 × volume of sinapinic acid matrix, and centrifuged at 13,000 rpm for 2 min

1 μL of supernatant was immediately added to its respective target well in triplicate