Figure 3.

TG-Induced Golgi Fragmentation Has Minor Effect on Protein Trafficking

(A) Cells were transfected with the Str-li_VSVG wt-SBP-EGFP plasmid for 16 h followed by a 30-min treatment with DMSO, 250 nM TG, or 10 μM monensin (Mo) at 37°C. Cells were then incubated with complete medium containing 40 μM biotin (chase) for the indicated times, lysed and treated with (+) or without (−) EndoH, and analyzed by Western blot for GFP.

(B) Quantification of (A) for the percentage of EndoH-resistant VSV-G from three independent experiments. Quantitation results are shown as Mean ± SEM. Statistical analyses were performed using two-tailed Student's t-tests by comparing with the control (∗p ≤ 0.05).

(C) Representative images of (A) showing the subcellular localization of VSVG-EGFP at indicated time points after biotin chase. Scale bar, 20 μm.

(D) Fluorescent images showing the subcellular localization of ManII-GFP in cells treated with DMSO (Ctrl) or 250 nM TG at the indicated time points of BFA washout. ManII-GFP appears in the Golgi area beginning at 60 min, whereas this takes longer in control cells. Scale bar, 20 μm.