Detection of LT and GAPDH was performed as previously described by Isoe J et al [20 (link)]. Guts from blood-fed female mosquitoes were homogenized in tissue extraction buffer (Invitrogen). Then, 10 μg protein was boiled in LDS (4×) NuPage sample buffer (Invitrogen) with 10× sample reducing agent (Invitrogen). For detection of LT, LT antibodies were used at a 1:200 dilution followed by the secondary anti-rabbit-HRP (Abcam) at a 1:5000 dilution. For detection of GAPDH, GAPDH monoclonal antibodies (Cell Signaling) were used at a 1:200 dilution followed by the secondary anti-mouse-HRP (Abcam) at a 1:5000 dilution.
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