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. 2024 Jan 9;15:380. doi: 10.1038/s41467-024-44696-3

Fig. 5. C. parvum utilizes stored glycogen (amylopectin) for glucose phosphate as a carbon source.

Fig. 5

a Schematic representation of stage I of glycolysis, glycogen breakdown and glycogen synthesis. b Schematic representation of Cp engineered to co-express the auxin receptor TIR1 from Oryza sativa and Cp glucan phosphorylase (CpGP) fused to mAID from Arabidopsis thaliana. See also Supplementary Fig. 1f for constructing and identification of CpGP-mAID-TIR1 transgenic parasites. c Schematic representation of conditional CpGP-mAID-TIR1 depletion. IAA, indole acetic acid; Ub, ubiquitin; SCF, Skp-1, Cullin, F-box (TIR1)-containing complex. d Immunofluorescence localization of CpGP-mAID-3HA expression. HCT-8 cells were infected with CpGP-mAID-TIR1 parasites and treated with 500 µM IAA or the vehicle (EtOH) for 24 h. Cells then were fixed and stained with rabbit anti-HA (green), mouse anti-TY (red), rat PanCp (cyan), and Hoechst (blue). The experiment was performed twice with similar outcomes. Scale bars = 2 µm. e Growth of CpGP-mAID-TIR1 parasites following treatment with IAA. HCT-8 cells were infected with CpGP-mAID-TIR1 parasites, treated with 500 µM IAA or the vehicle (EtOH) for 24 h or 48 h. Cells were fixed, and labeled with rabbit PanCp and Hoechst, and the number of Cp in each well was determined by using a Cytation 3 imager. Each bar represents the mean ± SD for nine replicates in total from three experiments. Statistical analysis performed using was performed by two-way ANOVA corrected for multiple comparisons by Sidak’s method. ns, not significant. ****, P < 0.0001. See also Supplementary Fig. 9c for the growth assay of host cell. f Transmission electron micrographs of CpGP-mAID-TIR1 parasites treated with vehicle (left) or IAA (right). HCT-8 cells were infected with CpGP-mAID-TIR1 parasites, and then treated with 500 µM IAA or the vehicle (EtOH). After 48 hpi, monolayers were fixed and processed for EM. Arrows (red) point to electron lucent amylopectin granules. Similar results were seen in multiple sections from one experiment. Scale bars = 500 nm. Source data are provided as an accompanying Source Data file.