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. 2015 Jan 7;32(4):1039–1055. doi: 10.1093/molbev/msu408

Fig. 1.

Fig. 1.

Isolation of the hydrogenosome-enriched fraction from Mastigamoeba balamuthi cell lysate. (A) Separation of the cell lysate (Lys) on a discontinuous OptiPrep gradient (15–40%) resulted in five distinct fractions (B1-5). The presence of double membrane-bound organelles in fraction B5 was confirmed using electron microscopy. (B) Immunoblot analysis of Lys and fractions B1-5 using polyclonal antibodies against the marker proteins mitHsp70, Cpn60, and MDH. (C) Distribution of marker enzymes (acid phosphatase for lysosomes and MDH for M. balamuthi hydrogenosomes) in fractions B1-5 after OptiPrep gradient centrifugation. (D) Immunofluorescence microscopy. Signal for mitochondrial marker protein Cpn60 (red) colocalized with hydrogenosomal MDH (green; PCC = 0.878 ± 0.036, n = 45).