Odyssey clx infrared imaging system software

Western blots were performed as previously described [55 (link)]. Briefly, ice-cold 0.04% saponin in 1x PBS was used to isolate parasites from host cells. Parasite pellets were subsequently solubilized in protein loading dye to which Beta-mercaptoethanol had been added (LI-COR Biosciences) and used for SDS-PAGE. Primary antibodies used in this study were rat-anti-HA 3F10 (Roche, 1:3000), mouse-anti-HA 6E2 (Cell Signaling Technology, 1:1000), rabbit-anti-HA 715500 (Thermofisher, 1:100), mouse-anti-V5 TCM5 (eBioscence, 1:1000), rabbit-anti-V5 D3H8Q (Cell Signaling Technology, 1:1000), rabbit anti-PfBiP MRA-1246 (BEI resources, 1:500), rabbit-anti-PfEF1α (from D. Goldberg, 1:2000), and mouse-anti-PfPMV (from D. Goldberg 1:400). Secondary antibodies used were IRDye 680CW goat-anti-rabbit IgG and IRDye 800CW goat-anti-mouse IgG (Li-COR Biosciences, 1:20,000). Membranes were imaged using the Odyssey Clx Li-COR infrared imaging system (Li-COR Biosciences). Images of membranes were processed using ImageStudio, the Odyssey Clx Li-COR infrared imaging system software (Li-COR Biosciences). Densitometry analysis of western blot signal was also performed using ImageStudio (Li-COR Biosciences).

Western blotting for Plasmodium parasites was performed as described previously (35 (link), 64 (link)). Briefly, parasites were selectively permeabilized by treatment with ice-cold 0.03% saponin in PBS for 15 min followed by lysis with RIPA to remove the hemozoin. The antibodies used in this study were mouse anti-HA antibody (clone 16B12, BioLegend; 1:500), rabbit polyclonal anti-HA SG77 (ThermoFisher; 1:500), mouse monoclonal anti-FLAG clone M2 (Sigma-Aldrich; 1:200), and mouse monoclonal anti-EXP2 clone 7.7 (1:500). The secondary antibodies used are IRDye 680CW goat anti-mouse IgG (LICOR Biosciences) (1:20,000). Western blot images were processed using the Odyssey Clx LICOR infrared imaging system software (LICOR Biosciences). Full-length blots are presented in Fig. S5.