Fig. 2: PfAnchor localizes to the cytoplasmic side of the apicoplast outer membrane.

a, b U-EXM analysis of PfAnchor localization in PfAnchor-iKD and PfAnchor^Mev-iKD cell lines. PfAnchor is detected in close proximity to non-nuclear DNA but does not colocalize with mitochondrial DNA. Apicoplast staining confirms that PfAnchor is specifically localized around the apicoplast. Representative images from one of three independent experiments performed on separate parasite cultures with similar results. Protein density in grayscale, PfAnchor (green), apicoplast (pink), mitochondria (yellow), and DNA (blue). Scale bars: 2 µm, with image depth (µm) indicated. Schematic representations highlight non-nuclear DNA (dark blue) and basal complexes (white), with light gray dashed lines indicating areas of potential membrane staining based on changes in NHS ester density. c Representative western blot from one of three independent experiments performed on separate parasite cultures. The proteinase K protection assay indicates that the C-terminal tagged PfAnchor is cytoplasmically exposed. Anti-PfAldolase and anti-GFP (PfACP) were used as unprotected and protected controls, respectively. Molecular weight markers are indicated on the left. Cropped regions are from the same blot. Full, uncropped scans are provided in the Supplementary Fig. 17.