Figure 2.
Figure 2. Colocalization of Sac2 with OCRL on early endosomes and at the last stage of clathrin-mediated endocytosis. Live spinning-disc confocal microscopy. (A) Colocalization of GFP-Sac2 and mCherry-OCRL. The majority of the fluorescent puncta positive for both proteins are expected to be early endosomes, given the close colocalization of both proteins with Rab5 (see Fig. 1 A and Hyvola et al., 2006; Erdmann et al., 2007). Insets show enlarged views of the regions boxed in yellow. (B and C) Time course of CLC-mRFP and either GFP-Sac2 (B) or GFP-OCRL (C) fluorescence intensities at very late stage clathrin-coated pits demonstrating that Sac2 and OCRL appear when clathrin is disappearing, and that Sac2 and OCRL display similar recruitment patterns. The disappearance of the Sac2 and OCRL fluorescence reflects the newly formed vesicle moving out of the focal plane. Bars: (A) 5 µm; (B and C) 1 µm.

Colocalization of Sac2 with OCRL on early endosomes and at the last stage of clathrin-mediated endocytosis. Live spinning-disc confocal microscopy. (A) Colocalization of GFP-Sac2 and mCherry-OCRL. The majority of the fluorescent puncta positive for both proteins are expected to be early endosomes, given the close colocalization of both proteins with Rab5 (see Fig. 1 A and Hyvola et al., 2006; Erdmann et al., 2007). Insets show enlarged views of the regions boxed in yellow. (B and C) Time course of CLC-mRFP and either GFP-Sac2 (B) or GFP-OCRL (C) fluorescence intensities at very late stage clathrin-coated pits demonstrating that Sac2 and OCRL appear when clathrin is disappearing, and that Sac2 and OCRL display similar recruitment patterns. The disappearance of the Sac2 and OCRL fluorescence reflects the newly formed vesicle moving out of the focal plane. Bars: (A) 5 µm; (B and C) 1 µm.

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