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. 2021 Aug 3;12(4):e01380-21. doi: 10.1128/mBio.01380-21

FIG 3.

FIG 3

TRAPP III-specific subunit TgTrs85 colocalized with TgStx5 is necessary for the transport of secreted proteins at the Golgi. (A) Immunofluorescence analysis of TgStx5 colocalization with TgTrs85. (B to E) Immunofluorescence analysis of the TgTrs85-mAID-3HA strain expressing the trans-, medial-, and cis-Golgi markers TgSORTLR-EGFP, TgAPμ1-EGFP, TgGalNac-3Myc, and TgGRASP-3Myc. (F) Immunofluorescence analysis of TgRab5A partially colocalized with TgTrs85. (G) The TRAPP core component TgBet5 partially colocalized with TgTrs85. The colocalization of TgTrs85 is shown, and signal intensity plots across the TgBet5 distribution are graphed. (H) Arrow in the left panel indicates the accumulation of residual bodies. The right panel shows the quantification of PVs with residual bodies in IAA-treated cultures and control. (I) Depletion of TgTrs85 led to defective transport of the secreted proteins. The left panel reveals the immunofluorescence staining of TgMIC2 in TgTrs85-mAID-3HA parasites cultured in the presence or absence of IAA for 48 h. The right panel shows quantification of parasites with diffused staining of TgMIC2 in IAA-treated cultures and the control. (J) Immunoblot analysis of microneme secretion. The left panel reveals conditional depletion of TgTrs85 affected secretion of TgAMA1 and TgMIC2. The tachyzoites were incubated for 24 h in the presence or absence of IAA. The right panel shows the semiquantitative analyses of Western blot signals of TgAMA1 and TgMIC2 in the ESAs and pellet.