Additional file 23: Additional Material 23-Supplementary Figure 9. Crude sub-cellular fractionation analysis of HA-GiCHMP7. HA-GiCHMP7-expressing transgenic trophozoites and non-transgenic control cells were subject to crude subcellular fractionation experiments through a freeze-thaw approach in liquid nitrogen to separate membrane-enriched and soluble fractions. Crudely fractionated protein samples, along with whole cell lysates from HA-GiCHMP7 transgenic lines and WB cells as a non-transgenic negative control, were subject to blotting for immuno-detection using an anti-HA antibody, followed by a secondary anti-Rat antibody coupled to HRP. A separate Coomassie gel was also prepared and depicted to demonstrate comparable amounts of protein were loaded for detection. The full experimental approach is reported in the Methods section. M: Marker; WB: non-transgenic total lysate; WCL: whole cell lysate; F-S: crudely-fractionated sample enriched for soluble components; F-M: crudely-fractionated sample enriched for membrane components.