Subcellular localisation of RhopH proteins during parasite invasion.

a Live RhopH2-mNeonGreen merozoites invading human erythrocytes imaged using lattice-light sheet microscopy. mNeonGreen signal (white arrows) visible as a bright spot on the merozoite apical end corresponding to RhopH2. The signal became diffused upon successful invasion as the parasitophorous vacuole formed. b Live RhopH3-mNeonGreen merozoites invading human erythrocytes imaged using lattice-light sheet microscopy. mNeonGreen signal (white arrows) visible as a bright spot on the merozoite apical end corresponding to RhopH3. The signal became diffused upon successful invasion as the parasitophorous vacuole formed. c Localisation of RhopH2-HA and RON4 in fixed merozoites at early, mid, late and complete stages of erythrocyte invasion. RON4 was a marker for the tight junction to assess the stage of invasion. Scale bar 2 µm. d Flag-RhopH3 and RON4 in fixed merozoites at early, mid, late and complete stages of erythrocyte invasion. RON4 was a marker for the tight junction to assess the stage of invasion. Scale bar 2 µm. e Clag3.1-HA and RON4 in fixed merozoites at early, mid, late and complete stages of erythrocyte invasion. RON4 was a marker for the tight junction to assess the stage of invasion. Scale bar 2 µm. f Super-resolution 3D reconstructions of invading merozoites. RON4 labelled in magenta marks the tight junction. HA-tagged Clag3.1, RhopH2, or RhopH3 are labelled in green. Clag3.1 signal appears in the host cytoplasm while RhopH2 and RhopH3 remain concentrated at the apical end of the invading merozoite. Pasternak M, Verhoef JMJ, Wong W, Triglia T, Mlodzianoski MJ, Geoghegan N, Evelyn C, Wardak AZ, Rogers K, Cowman AF. RhopH2 and RhopH3 export enables assembly of the RhopH complex on P. falciparum-infected erythrocyte membranes. Commun Biol. 2022 5(1):333. PMID: 35393572