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Fig. 5 | BMC Biology

Fig. 5

From: The divergent ER-mitochondria encounter structures (ERMES) are conserved in parabasalids but lost in several anaerobic lineages with hydrogenosomes

Fig. 5

Western blot analysis of T. vaginalis ERMES components in subcellular fractions. T. vaginalis cells expressing A TvMmm1a, B TvMdm12, C TvMmm2a, or TvMmm2b were sonicated and subcellular fractions were separated using differential and Percoll gradient centrifugation. L, cell lysate; C, cytosolic fraction; LDV, low-density vesicles; H, hydrogenosomal fraction. TvMmm1a, TvMmm2a/b, and TvMdm12 were detected using mouse α-HA antibodies. Cyt. ME, cytosolic malic enzyme (cytosolic marker); OsmC (hydrogenosomal marker); and PDI (ER marker) were visualized by mouse α-cytosolic ME, rat α-OsmC, and rat α-PDI polyclonal antibodies, respectively. D TvMmm2 topology test. TvMmm2a and TvMmm2b hydrogenosomal fraction (H) was treated with proteinase K (HpK) and proteinase K with Triton X-100 (HpKTX). TvMmm2a and TvMmm2b were detected by mouse α-HA antibody, OsmC was detected by rat polyclonal α- OsmC antibody, and C-tail anchored protein 7 (CTA7, outer hydrogenosomal membrane marker protein) was detected by rat polyclonal α-CTA7 antibody

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BMC Biology

ISSN: 1741-7007

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