Figure 2-.

TgGC is essential for the lytic life cycle of T. gondii (A) Domain architecture of TgGC showing functional (NCBI CDD) and transmembrane domains (OCTOPUS) as well as insertion points for N- and C-terminal tags. See also Figure S2A. (B) IF microscopy of intracellular RH 6Ty-GC-mAID-3HA parasites treated with indole acetic acid (IAA) or vehicle for 14 h. Parasites were labeled with mouse anti-Ty and anti-mouse IgG Alexa Fluor 488 (green), rat anti-HA and anti-rat IgG Alexa Fluor 568 (red), and rabbit anti-TgGAP45 and anti-rabbit IgG Alexa Fluor 647 (magenta). Scale bar = 5 μm. See also Figure S2B. (C) Immunoblot of lysates from RH TIR1-3FLAG and derivative RH 6Ty-GC-mAID-3HA parasites treated with IAA or vehicle for 14 h. Blots were probed with rabbit anti-Ty and anti-rabbit IgG IRDye 680RD and mouse anti-HA and anti-mouse IgG IRDye 800CW. Separate channels of the same membrane scan are shown. See also Figure S2C. (D-E) Plaques formed by TIR1-3FLAG and derivative TgGC tagged lines on HFF monolayers with IAA or vehicle control (Ctrl) at D8 with 200 parasites/ monolayer. Scale bar = 5 mm. (E) Plaque data represented as mean ± SD (n = 9 replicates combined from N = 3 trials). Each parasite line was analyzed individually for statistical significance using an unpaired t test (IAA vs control), P values: **** ≤ 0.0001.