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

Figure 4.

Figure 4

ESCRT-III Proteins and VPS4A Concentrate at Flemming bodies. (A) Double labeled immunofluorescence images showing that CHMP2A-FLAG, CHMP4A-FLAG, and CHMP5-FLAG (each 0.5 μg DNA) concentrate in double ring structures (arrowheads) at the midbodies of dividing HeLa cells. Expanded views are shown below each image. Note that these ESCRT-III constructs were employed because they exhibited no (CHMP2A-FLAG and CHMP4A-FLAG) or minimal (CHMP5-FLAG) effects on HIV budding (von Schwedler et al, 2003). Microtubules were also stained for reference (red, α-Tubulin). (B) Double-labeled immunofluorescence images showing that endogenous VPS4A concentrates in double ring structures (arrowheads) within very thin midbodies of dividing HeLa cells. Microtubules were also stained (red, α-Tubulin). Examples of wider midbodies that lacked VPS4A staining are shown in Supplementary Figure S8A, and VPS4 localization is quantified in Supplementary Figure S8B.