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. 2010 Sep 20;190(6):1005–1022. doi: 10.1083/jcb.200912089

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© 2010 Mari et al.

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Figure 5. — Atg9 concentrates into a novel organelle. (A) Atg9-GFP colocalizes with some FM 4-64-positive puncta shortly after endocytosis. The strains expressing genomically GFP-tagged Tlg1 (FRY360) or Atg9 (FRY162) were grown to log phase and then exposed to FM 4-64 as described in Materials and methods. Arrowheads highlight the colocalization between FM 4-64 and GFP-Tlg1 or Atg9-GFP. (B) Atg9 does not colocalize with endosomal and Golgi protein markers. The Atg9-GFP Vrg4-mChe-V5 (FRY340), Atg9-GFP Sec7-dsRED (FRY341), GFP-Tlg1 Atg9-mChe-V5 (FRY342), and GFP-Pep12 Atg9-mChe-V5 (FRY344) strains were grown to log phase before being fixed and imaged. Bars, 2 µm. (C) The colocalization experiments shown in B were statistically evaluated as described in Materials and methods. Atg23 was used as a positive control for colocalization because forming a complex with Atg9 (Reggiori et al., 2004a). DIC, differential interference contrast.