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. 2018 Mar 1;8:53. doi: 10.3389/fcimb.2018.00053

Figure 2.

Figure 2

ESCRT-III proteins are located in cytoplasm, vesicles, nuclei and around phagosomes of trophozoites. (A) Coomassie blue stained polyacrylamide gel of the recombinant proteins (without GST-tag). (B) Western blot of trophozoites lysates (TL) and purified GST-tagged recombinant proteins (R) using specific antibodies or pre-immune sera (PS). (C) Representative images of 4% paraformaldehyde fixed trophozoites treated with the anti-ESCRT-III proteins antibodies (green) in basal conditions (0 min) and during erythrophagocytosis. Arrowheads show the antibodies signal in erythrophagosomes. Arrows shows the presence of the proteins in the nuclei stained by DAPI. PS: pre-immune serum. (D) Open field of trophozoites stained by α-rEhVps24 or α-rEhVps2 and corresponding secondary antibodies and DAPI after 60 min of erythrophagocytosis. (E) Representative images of 4% paraformaldehyde fixed trophozoites treated with the αEhVps24 or α-rEhVps2 (green) and α-pCNA protein (red) antibodies after 60 min of erythrophagocytosis. Nuclei were stained by DAPI. (F) Open field of trophozoites stained by αEhVps24 or α-rEhVps2 and α-pCNA with corresponding secondary antibodies after 60 min of erythrophagocytosis. phc, phase contrast. Scale bar: 10 μm.