Tissue RNA-seq expression data and results of venom LC-MS analysis for Hylyphantes graminicola

The protein samples were then concentrated (20 min at 80 V) and separated (90 min at 120 V) in 12% SDS-PAGE gel electrophoresis. The gel was stained by Coomassie Brilliant Blue R-250 and decolored until the bands were visualized clearly. Trypsin treatment was performed at 37°C for 4 h.

Peptides were washed three times in 0.1% formic acid (FA) and 3% acetonitrile (ACN) and once in 0.1% FA and 70% ACN. The lyophilized samples were then dissolved in solution A (100% water and 0.1% FA) and injected into a homemade C18 Nano-Trap column (4.5 cm×75 μm, 3 μm) at 55°C. The separated peptides were analyzed by Orbitrap Exploris 480 matched with FAIMS (Thermo Fisher), with an ion source of Nanospray Flex™ (ESI), an ion transport capillary temperature of 320°C, and a spray voltage of 2.1 kV.

The compensation voltage of FAIMS was set as -45 and -65, with a scan round time of 1 s, a resolution of 15,000, and an automatic gain control (AGC) target value of 750,000.