1. The trafficking of VSVG–GFP to pre‐Golgi structures is impaired by depletion of GMAP‐210.The trafficking of VSVG–GFP to pre‐Golgi structures is impaired by depletion of GMAP‐210.
The trafficking of VSVG–GFP to pre‐Golgi structures is impaired by depletion of GMAP‐210.The trafficking of VSVG–GFP to pre‐Golgi structures is impaired by depletion of GMAP‐210. (A) Luciferase (siLuc) or GMAP‐210 (siGMAP #2)‐depleted HeLa cells were infected with ts045‐VSVG–GFP and incubated at 40°C for 16 h. Cells were shifted to 17°C for 1 h to induce VSVG–GFP accumulation in the ERGIC prior to staining for endogenous ERGIC53. (B) Quantification of VSVG–GFP localization (50–100 cells per condition) from A. Data show mean±s.e.m from three independent experiments. *P<0.05 (paired Student's t‐test). (C) HeLa cells transfected with the indicated siRNAs were infected with ts045‐VSVG–GFP adenovirus, and incubated at 40°C for 16 h. Cells were then placed on ice for 15 min with nocodazole (NCZ), and either fixed directly, or warmed to 31°C in the presence of the drug for 20 min before staining for endogenous GalNAcT2. (D) HeLa cells expressing GMAP‐210–Myc and ts045‐VSVG–GFP were placed on ice for 15 min with NCZ. Cells were shifted to 31°C for 20 min in the presence of NCZ and then stained for (upper panel) the Myc epitope (red) or (lower panel) the Myc epitope (green) and endogenous ERGIC53 (red). Scale bars: 10 µm.
Peristera Roboti et al. J Cell Sci 2015;128:
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