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cmi12132-sup-0001-fs1.jpg1324K

Fig. S1. Sections from ROTO prepared tomograms of free intact Plasmodium falciparum merozoites.

A–C. Virtual sections through a merozoite showing connection between the inner membrane complex (IMC) and Endoplasmic reticulum (ER) membranes. Each image represents a 10 nm slice and each image is separated by 15 nm.

D–F. Virtual sections through a merozoite showing connection between the inner membrane complex (IMC) and Endoplasmic reticulum (ER) membranes. Scale bars are 200 nm.

cmi12132-sup-0002-fs2.jpg1173K

Fig. S2. Cryo-electron tomography of a free intact Plasmodium falciparum merozoite.

A. Tomogram of a free intact Plasmodium falciparum merozoite. Note the discontinuities in the inner membrane complex (arrowheads).

B. Model constructed from CET of the apex of a free merozoite showing the distinct interruptions behind the apical third of the inner membrane complex (green). Rhoptries are shown in pink, apical rings are shown in yellow, microtubules are shown in dark blue, plasma membrane is shown light blue. Scale bars are 200 nm.

cmi12132-sup-0003-fs3.jpg1113K

Fig. S3. Immunoelectron microscopy (IEM) of merozoites pre- and post-erythrocyte invasion. After sectioning, parasites are labelled with primary antibody then primary antibody is detected with a secondary antibody conjugated to colloidal gold (white arrows). Scale bars are 200 nm.

A. IEM of free merozoite (pre-invasion) labelled with the microneme (Mi) marker anti-AMA1.

B. IEM of free merozoite (pre-invasion) labelled with the rhoptry (Rh) maker anti-RAP1. IEM of merozoite (post-invasion) labelled with dense granule (Dg) marker anti-RESA.

cmi12132-sup-0004-movie 1.mov824K

Movie S1. Electron tomography of a whole cryo-preserved Plasmodium falciparum merozoite. Cryo-electron tomogram of a free, intact Plasmodium falciparum merozoite. The tomogram was acquired on a FEI Titan Krios, and reconstructed with IMOD. Two rhoptries, numerous micronemes and two microtubules are visible at the apex of the merozoite. Note the opening in the inner membrane complex starting at the third of the apical rings.

cmi12132-sup-0005-fsmovie 2.mov2075K

Movie S2. Serial electron tomogram of a ROTO-fixed free merozoite. Tomogram of a free Plasmodium falciparum merozoite reconstructed from serial sections. The tomogram was acquired on a Tecnai F30 (FEI, NL), reconstructed and segmented with IMOD. In the model, the nucleus is represented in red, the apicoplast in blue, the mitochondria in orange, the rhoptries in cyan and the dense granule and microneme are multicoloured. The continuum of the IMC/nuclear membrane and the ER is represented in gold.

cmi12132-sup-0006-movie 3.mov1030K

Movie S3. Serial electron tomogram of a GF-fixed merozoite with fusing rhoptries. Tomogram of the apex of an invading P. falciparum merozoite reconstructed from serial sections. The tomogram was acquired on a Tecnai F30 (FEI, NL), reconstructed with IMOD. The tomogram span 3 physical sections and details the entire volume of fusing rhoptries from the neck to the top of the bulbs.

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