Mouse anti c myc 9b11

Parasite extracts were resuspended in 2× Laemmli sample buffer (Bio-Rad) with 5% 2-mercaptoethanol (Sigma-Aldrich). Samples were boiled for 5 min at 95°C before separation on a gradient 4–20% SDS-PAGE gels (Bio-Rad). Samples were then transferred to nitrocellulose membrane using standard methods for semidry transfer (Bio-Rad). Membranes were probed with rabbit anti-HA (CS29F4 cat. no. 3724S, Cell Signaling Technology), mouse anti-c-Myc (9B11 cat. no. 2276, Cell Signaling Technology) or mouse anti-F₁B ATPase (made in house) all at a dilution of 1:5000, or rat anti-IMC10 (made in house) at a dilution 1:5000 overnight. Given the high molecular mass of ATPase-guanylyl cyclase, we used 4–20% Tris-acetate SDS gels (Invitrogen) as performed in Yang et al. (2019b) (link). Membranes were then washed and probed with either goat anti-mouse-IgG horseradish peroxidase or goat anti-rabbit-IgG horseradish peroxidase (Sigma-Aldrich) at a dilution of 1:10,000 for 1 h (GE Healthcare). Proteins were detected using SuperSignal West Femto substrate (Thermo Fisher) and imaged using the FluorChem R system (Biotechne). Full original western blots are shown in Fig. S6.