Protein Extraction and Western Blot Analysis of Locust Muscle

Total proteins from flight muscles of WT and ACP^-/- locusts were extracted using TRIzol reagent, as previously described (Hou et al., 2019 (link)). The protein extracts (50 μg) were electrophoresed on 4–20% Biofuraw precast gels (Tanon, China) and then transferred to polyvinylidene difluoride (PVDF) membranes (Millipore). The membrane was incubated with polyclonal antibody against target protein (anti-FABP, developed by ABclone, China, 1:5000; anti-CPT2, Abcam, 1:1000; anti-ACDM, Abcam, 1:1000). Goat anti-rabbit IgG (EASYBIO, 1:5000) was used as the secondary antibody. Polyclonal antibody against tubulin was used as an internal control. Protein bands were detected by chemiluminescence (ECL kit, Thermo Scientific).