Fig. 1

NcGRA17-BirA* localizes to the PV and can biotinylate proteins in the vacuole. a Diagram of the construct encoding the full genomic sequence of NcGRA17 fused to BirA* along with a 3 × HA C-terminal epitope tag. b IFA showing that NcGRA17-BirA* traffics appropriately to the PV and co-localizes with NcGRA7 (scale bar = 5 µm). c IFA of NcGRA17-BirA* expressing parasites ± biotin, showing the PV is labeled in a biotin-dependent manner. Endogenously biotinylated apicoplasts are observed with biotin (arrowheads) (scale bar = 5 µm). d Western blotting of whole-cell lysates of parent (Nc1) and NcGRA17-BirA*-expressing parasites ± biotin. Lysates were probed with streptavidin-HRP, revealing an increase in biotinylated proteins in NcGRA17-BirA*-expressing parasites upon the addition of biotin