Schizont-processed Rh1-240 colocalizes with Rh1-120 at

the apical tip and tight junction of the merozoite.

A. The Rh1-120 protein is located at the apical tip and tight junction

of merozoites. Free merozoites from the W2mef-Rh1HA line were

dual stained with a McAb to RAP1 in the body of the rhoptry and

with rat anti-HA antibodies to the tagged 120 kDa Rh1. Anti-HA

staining is seen either apical to the RAP1 staining (top panels) or

at both sides but posterior to RAP1 staining indicating tight junction

staining (bottom panels). Nuclei were stained with DAPI.

B. The Rh1-120 protein colocalizes with the Rh1-240 protein at the

apical tip and tight junction of merozoites. Free merozoites from the

W2mef-Rh1HA line were dual-stained with rat anti-HA antibodies to

the tagged 120 kDa Rh1 and with a rabbit antibody (R515) to the

240 kDa Rh1. Colocalization is seen at the apical tip (panels a–d)

and at the tight junction (panels e–l) of individual merozoites.

C. The Rh1-240 protein colocalizes with EBA-175 at the tight

junction of merozoites. Free merozoites from the W2mef-175His

line were dual-stained with mouse anti-His antibodies to tagged

EBA-175 and with a rabbit antibody (R515) to the 240 kDa Rh1.

EBA-175 and Rh1-240 appear to colocalize at the tight junction.

D. Free merozoites from the W2mef-Rh1HA line were dual-stained

with rat anti-HA antibodies to the tagged 120 kDa Rh1 and with a

rabbit antibody (R515) to the 240 kDa Rh1. Simultaneous staining

of both the apical tip and the tight junction of a merozoite is seen.

E–G. Merozoites of the W2mef-Rh1HA or the W2mef-175His lines

were trapped during invasion using cytochalasin D. Rh1-240 was

stained with R515, Rh1-120 with rat anti-HA and EBA-175 with

mouse anti-His antibodies. PfRON4 was stained either with a

mouse antibody (E) or with a rabbit antibody (F and G). Evers F, Roverts R, Boshoven C, Kea-Te Lindert M, Verhoef JMJ, Sommerdijk N, Sinden RE, Akiva A, Kooij TWA. Comparative 3D ultrastructure of Plasmodium falciparum gametocytes. Nat Commun.