Confocal microscopy demonstrates patching of different ETRAMPs into distinct domains in formaldehyde-fixed IRBCs. IRBCs (D10E2myc parasites) coated on 10-well slides using concanavalinA (fixed with 1% formaldehyde and permeabilized with 0.03% saponin) were incubated with sera specific for ETRAMP2, ETRAMP10.1, ETRAMP2myc (detected via the tag) or anti-ETRAMP2 serum labelled with a fluorescent Fab fragment and viewed by confocal microscopy. A. ETRAMP2 (cy2, green) co-patches with ETRAMP2myc (cy3, red) into distinct areas in the PVM of several ring-stage IRBCs. B. ETRAMP10.1 (cy2, green) patches into distinct areas when compared with ETRAMP2myc (cy3, red) in several ring-stage IRBCs. bf, brightfield; bar size, 4 μm.Spielmann T, Gardiner DL, Beck HP, Trenholme KR, Kemp DJ. Organization of ETRAMPs and EXP-1 at the parasite-host cell interface of malaria parasites. Mol Microbiol. 2006 59:779-94. PMID:

Colocalization of ETRAMPs with EXP-1 and SERP. The first image in each set shows ETRAMP labeling (green, FITC), the second Texas-Red-labeled SERP (A) or EXP-1 (B–F), and the third image represents the merged picture. The fourth panel in B and C depicts the merged picture, including nuclear staining (DAPI). (A) Confocal microscopy analysis of SERP and ETRAMP4 colocalization in schizont stage parasites (dried and 1% formaldehyde fixed). The ETRAMP4 signal seems partially detached from the PV content (arrow). (B and C) ETRAMP4 and EXP-1 localize to dots, probably representing vesicular structures that are detached from the PVM (E) or seem connected to the PVM (F). Dots are indicated by arrows. (D and E) Fluorescent microscopic analysis of ETRAMP and EXP-1 costaining of a ring-stage parasite displaying an even circular pattern (D, 0.1% formaldehyde-fixed IRBCs, ETRAMP10.1) or a beads on a string pattern (E, unfixed IRBCs, ETRAMP2). Yellow color shows regions of overlap (arrows). This indicated that ETRAMPs and EXP-1 localize to different regions of the PVM.Spielmann T, Fergusen DJ, Beck HP. etramps, a new Plasmodium falciparum gene family coding for developmentally regulated and highly charged membrane proteins located at the parasite-host cell interface. Mol Biol Cell. 2003 14:1529-44.