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[Preprint]. 2026 May 24:2026.05.21.726867. [Version 1] doi: 10.64898/2026.05.21.726867

Figure 5: Loss of TGIF affects newly synthesized MIC2 trafficking.

Figure 5:

(A) Left. Schematic of the pulse-chase recycling assay. Right. Immunofluorescence showing recycled MIC2 stained with Halo-TMR (green) in TGIF-Ty:MIC2-Halo parasites grown for 24 hours with ethanol (control) or IAA (knockdown). Scale bars are 5μm. Representative images from three experimental replicates (N> 45 vacuoles per condition). (B) Schematic of the pulse-chase trafficking assay. (C) TGIF-Ty:MIC2-Halo parasites labeled with HaloLigand-TMR (green) and HaloLigand-JF646 (magenta) after growth with ethanol (control) or IAA (knockdown). Time points indicate period of growth between 1st (TMR) and 2nd (JF646) labeling reactions. Scale bars are 5μm. Representative images from three experimental replicates (N > 300 vacuoles per time point per condition). (D) Stacked bar chart indicating the localization of MIC2 labeled with JF646 at each time point. P-values are indicated. (E) Bar graph indicating the percent of vacuoles with JF646 staining at each time point, indicating the presence of newly synthesized protein. P-value indicated. (E) Illustration of MIC2 trafficking pathway in control and TGIF knockdown parasites.