FIG 1 .

Lectin profiling of ppGalNAc-T-knockout strains. Purified lysates from T. gondii strains were separated by SDS-PAGE and transferred onto a PVDF membrane which was then incubated with either biotin-conjugated jacalin (A) or VVA (B). The orange arrowheads indicate the bands missing in both ppGalNAc-Δt2 and -Δt3 strains. The blue arrowheads indicate the bands missing in ppGalNAc-Δt2 but not in -Δt3. The gray arrowheads indicate the nonspecific endogenous biotinylated proteins (streptavidin was used for detection). This blot demonstrates that multiple T. gondii glycoproteins require ppGalNAc-T2 and -T3 glycosyltransferase activities. The duplicate blots on the right were subjected to β-elimination and demonstrate that the identified bands are due to O-linked glycosylation. This figure demonstrates T. gondii cultivated under tachyzoite (pH 7.1 with 5% CO₂) culture conditions, and Fig. S3 in the supplemental material demonstrates T. gondii cultivated under bradyzoite culture conditions (pH 8.1 with 0.1% CO₂). WT, wild type.