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. Author manuscript; available in PMC: 2012 Aug 23.
Published in final edited form as: Nature. 2012 Feb 22;482(7386):495–500. doi: 10.1038/nature10822

Figure 4. Cul3Klhl12-dependent monoubiquitination enlarges COPII-structures.

Figure 4

a. Localization of induced FLAGKlhl12 (green) and Sec31 (red) in 293T cells, monitored by confocal microscopy. Scale bar: 3μm. b. Klhl12-expressing HeLa cells were analyzed for Klhl12 (green) and Sec31, Sec13, or Sec24C (red) by confocal microscopy. Scale bar: 3μm. c. COPII-structures in HeLa cells transfected with FLAGKlhl12, lysine-free ubiquitin, or Cul3-siRNA, analyzed by confocal microscopy. Scale bar: 500nm. d. Upper panel: Thin-section EM of Klhl12-expressing or control HeLa cells (red arrow: Klhl12-dependent structures; blue arrow: small control vesicles). Scale bar: 500nm. Lower panel: Immunogold-EM of Klhl12 in transiently transfected HeLa (left) or stable 293T cells (right). Scale bar: 200nm. e. HeLa cells transfected with FLAGKlhl12, lysine-free ubiquitin, FLAGKlhl12FG289AA, FLAGCul31-250, or FLAGCul3, were analyzed for localization of Klhl12/Cul3 (green) and Sec31 (red) by confocal microscopy. Scale bar: 5μm.