FIGURE 8.

TUG depletion alters GM130 staining intensity and impairs Golgi reformation after brefeldin A washout. A, endogenous TUG and GM130 were imaged using immunofluorescence and confocal microscopy of HeLa cells. Cells were treated with control siRNA, TUG siRNA A, or TUG siRNA B, as indicated, for 48 h prior to fixation. B, HeLa cells were treated with control siRNA or TUG siRNA A for 48 h and then mock-treated or treated with BFA, as indicated. BFA was used at 10 μg/ml at 37 °C for 1 h, and then cells were washed three times and allowed to recover in the absence of BFA for 1 h at 37 °C. Cells were then fixed and stained to detect GM130 as in A. The experiment was repeated with similar results. C, images of cells in B were quantified. For each cell, the ratio of fluorescence intensity in the perinuclear region to that of the entire cell was quantified. 19–24 cells were analyzed per condition, and the mean ratio ± S.E. (error bars) is plotted. Statistical significance was assessed using a two-tailed t test. Scale bars, 20 μm.