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. 2015 Apr 10;4(5):636–648. doi: 10.1242/bio.201511551

Fig. 2. Clu localizes with dGRASP and Sec16 ER exit sites.

Fig. 2.

(A–G) Muscle tissue from L3 larvae was dissected and immunostained to examine the subcellular localization of Clu protein. (A,A′) The Clu:GFP protein trap line (green) localizes in a repeated pattern within the muscle (arrows) and accumulates around the nuclei (n; indented arrows). The right panel is a close up of the boxed region in the left panel. (B) The perinuclear accumulation of Clu:GFP (green) reveals little colocalization with mitochondria (red; anti-Complex V). (C–F) An anti-Clu antibody (red) was used to confirm the Clu:GFP nuclear staining pattern and also to discern the localization of Clu puncta with other organelle markers (green) in WT larval muscle. A composite Z-stack is followed by representative single confocal slices. (C,D) Clu-positive puncta overlap with both a general ER marker (C) and the ERES protein Sec16 (D). (E,F) Clu colocalizes with a subset of Golgi:YFP puncta (E). (F) The percentage of puncta of each organelle marker that overlap with Clu protein. Colocalization was determined from multiple single plane images calculated using the Image J JACoP plug-in. Mean±s.e.m. Scale bars, 50 µm (A), 10 µm (A′,B–G).