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

Figure 4: Microneme protein localization and processing is affected by the loss of TGIF.

Figure 4:

(A) Immunofluorescence showing the localization of microneme proteins proM2AP (anti-proM2AP, magenta, top row), M2AP (anti-M2AP, magenta, middle row), and MIC2 (anti-MIC2, magenta, bottom row) in TGIF-Ty parasites grown with ethanol (control) or IAA (knockdown) for 24 hours. Parasite periphery is indicated using anti-MLC1 antibodies (yellow). White arrowhead denotes the localization of proteins at the Golgi or post-Golgi compartments. White arrow shows protein localized at mature micronemes. Scale bars are 5μm. Representative images from two experimental replicates (N = 30 vacuoles per condition). (B) Immunofluorescence showing the microneme proteins AMA1 (anti-AMA1, green) and MIC8-mCherry (magenta) grown in TGIF-Ty parasites with ethanol (control) and IAA (knockdown) for 24 hours. White arrowhead denotes the localization of proteins at the Golgi or post-Golgi compartments. White arrow protein localization at mature micronemes. Scale bars are 5μm. Representative images from two experimental replicates (N = 20 vacuoles per condition). (C) Western blot with anti-proM2AP, anti=M2AP, anti-MIC2, and anti-tubulin (loading control) on TGIF-Ty parasite lysates harvested after 36 hours of ethanol or IAA treatment. Representative blots from three experimental replicates. (D) Immunofluorescence showing ROP1 fused to neonFP (magenta; top) grown in TGIF-Ty parasites with ethanol (control) or IAA (knockdown) for 24 hours. Parasite periphery is indicated using anti-MLC1 antibodies (yellow; bottom). Scale bars are 5μm. Representative images from three experimental replicates (N > 40 vacuoles per condition). (E) Western blot with anti-ROP1 and anti-IMC1 (loading control) on TGIF-Ty parasite lysates harvested after 36 hours of ethanol and IAA treatment. Representative blots from three experimental replicates.