Western Blot Analysis of rSjVAMP2 Protein

At 6 days post-siRNA treatment, soluble proteins were extracted from worms and quantified by using the Bradford protein assay (Sangon Biotech, Shanghai, China). Subsequently, the proteins were separated by SDS-PAGE, electrotransferred onto a PVDF membrane (Whatman International Ltd., Kent, UK), and blocked in 5% skim milk in PBS (pH 7.4) containing 0.1% Tween 20 (Sigma, St Louis, MO, USA) (PBST) overnight at 4 °C. The membrane was incubated with specific mouse anti-rSjVAMP2 serum^{43 (link)} at a dilution of 1:100 or β-actin (Cell Signaling Technology, Boston, MA, USA) diluted 1:1000 in PBST for 1 h at room temperature. After three 5-min washes in PBST, the bound primary antibodies were detected using horseradish peroxidase-conjugated goat anti-mouse IgG (Beyotime Biotechnology, Shanghai, China) diluted 1:2000 for 1 h at room temperature. The membrane-bound proteins were exposed to an X-ray film and detected using the enhanced chemiluminescence detection system (GE Healthcare Life Sciences, Stockholm, Sweden).