Figure 6. ES_I delays anterograde trafficking of tsO45 VSV-G-GFP.

A. HeLa cells infected with adenovirus encoding tsO45 VSV-G-GFP were grown overnight at 40°C, then pre-treated with DMSO, with 8 µM ES_R35, ES_I, or with 5 µg.ml^-1 Brefeldin A (BFA) for 1 h. Cells were then shifted to 32°C, and incubated for the time indicated in the continued presence of the compounds, prior to fixation and visualisation by fluorescence microscopy. B. Lysates of HeLa cells treated as in A were incubated with Endoglycosidase H (EndoH, +) or mock treated (−), then analysed by SDS-PAGE followed by immunoblotting using anti-VSVG antibody and alkaline phosphatase conjugated secondary antibody. The position of N-glycosylated (CHO) and non-glycosylated VSV-G are indicated.