Supplementary Figure 2: Vps54 and Syndetin exhibit distinct organelle targeting and retrograde transport activities.
From: EARP is a multisubunit tethering complex involved in endocytic recycling
(a) Co-localization of Vps54-EGFP and Syndetin-EGFP with Ang2–13Myc in rat hippocampal neurons analyzed with antibodies to GFP, the myc epitope and MAP2 as described in the legend to Figure 3. Bars, 10 μm. (b) Syndetin KD does not affect retrograde transport from endosomes to the TGN. Control (mock), Ang2-, Vps54- and Syndetin-KD HeLa cells were treated with 0.5 μg/ml Cy3-conjugated B-subunit of Shiga toxin (STxB) at 37 °C for 15 min and chased for 1 h before fixation and immunostaining. Nocodazole treatment was applied at 10 μM for 2 h following the chase where indicated. Giantin (medial Golgi marker) and TGN46 (TGN marker) were visualized by immunostaining with rabbit antibody to Giantin and sheep antibody to human TGN46 followed by Alexa647 donkey anti-rabbit and Alexa488 donkey anti-sheep antibodies. Bars, 10 μm.
