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. 2007 Sep 13;26(19):4215–4227. doi: 10.1038/sj.emboj.7601850

Figure 3.

Figure 3

CEP55, ALIX, and TSG101/ESCRT-I colocalize at centrosomes and Flemming bodies. (A) Triple labeled immunofluorescence and DIC images showing that FLAG-CEP55 (0.2 μg DNA), GFP-TSG101/ESCRT-I (0.2 μg GFP-TSG101, VPS28, VPS37B, and MVB12A DNA), or GFP-ALIX (0.5 μg DNA) colocalize at the midbodies (arrowheads) of dividing HeLa cells. Microtubule staining (white, α-Tubulin) is also shown for reference in columns 1 and 5. Expanded and merged views of the midbodies from the lower pair of cells in each image are shown in column 5. (B) Double labeled immunofluorescence and DIC images showing that α-Tubulin, FLAG-CEP55, or CEP55-Myc (0.2 or 0.5 μg DNA), Myc-TSG101/ESCRT-I (0.2 μg Myc-TSG101, VPS28, VPS37B, and MVB12A DNA), and FLAG-ALIX (0.5 μg DNA) colocalize at the centrosomes of non-dividing cells (arrowheads). (C) Triple-labeled immunofluorescence and DIC images showing that GFP-CEP55 (0.5 μg DNA) localizes to midbodies (arrowheads) whereas OSF-TSG101/ESCRT-I (0.2 μg GFP-TSG101154–164A, VPS28, VPS37B, and MVB12A DNA) and FLAG-ALIX800-802A (0.5 μg DNA) proteins that cannot bind CEP55 are not recruited to midbodies. Microtubule staining (white, α-Tubulin) is also shown for reference.