Figure 3.
Figure 3. VPS28 binds ESCRT-II and CHMP6 to recruit CHMP4B. (A) Schematic view of the VPS28 constructs used. (B) Confocal images of HeLa cells stably expressing siRNA-resistant VPS28 alleles and mCherry-TSG101 upon siRNA treatment as indicated, stained for DNA (Hoechst), α-tubulin, and mCherry. Bars, 5 µm. (C) Confocal images of CHMP6 recruitment in HeLa cells stably expressing VPS28 alleles. Bars, 5 µm. (D) Schematic view of cytokinetic CHMP4B recruitment pathways. (E) Confocal images of CHMP4B recruitment in HeLa cells stably expressing VPS28 alleles. Bars, 5 µm. (F) Quantification of CHMP4B at the midbody (error bars indicate SEM; n ≥ 23 cells from four independent experiments; unpaired t test; ***, P < 0.001). Knockdown of ALIX and expression of siRNA-resistant VPS28 is shown by Western blot.

VPS28 binds ESCRT-II and CHMP6 to recruit CHMP4B. (A) Schematic view of the VPS28 constructs used. (B) Confocal images of HeLa cells stably expressing siRNA-resistant VPS28 alleles and mCherry-TSG101 upon siRNA treatment as indicated, stained for DNA (Hoechst), α-tubulin, and mCherry. Bars, 5 µm. (C) Confocal images of CHMP6 recruitment in HeLa cells stably expressing VPS28 alleles. Bars, 5 µm. (D) Schematic view of cytokinetic CHMP4B recruitment pathways. (E) Confocal images of CHMP4B recruitment in HeLa cells stably expressing VPS28 alleles. Bars, 5 µm. (F) Quantification of CHMP4B at the midbody (error bars indicate SEM; n ≥ 23 cells from four independent experiments; unpaired t test; ***, P < 0.001). Knockdown of ALIX and expression of siRNA-resistant VPS28 is shown by Western blot.

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