Effect of Golgi disrupting treatments on the Golgi apparatus of MDA-MB-231 cells.

<p>Cells were left untreated (A; <i>Control</i>), or transfected to transiently express the HA-epitope-tagged ARF1 constitutively-activated mutant for 16 h (B; <i>HA-ARF1-Q71L</i>), or treated for 60 min either with 5 μg/ml Brefeldin A (C; <i>BFA</i>) or 10 μM Golgicide A (D; <i>GCA</i>). Cells were fixed, permeabilized, and immunolabeled with mouse monoclonal antibody to GM130, rabbit polyclonal antibody to Giantin, and sheep antibody to TGN46. Secondary antibodies were Alexa-594-conjugated donkey anti-mouse IgG (red channel), Alexa-488-conjugated donkey anti-rabbit IgG (green channel), and Alexa-647-conjugated donkey anti-sheep IgG (blue channel). Nuclei were stained with DAPI (gray channel). Stained cells were examined by fluorescence microscopy. Merging red, green, blue, and grey channels generated the fourth image on each row; yellow indicates overlapping localization of the red and green channels, cyan indicates overlapping localization of the green and blue channels, magenta indicates overlapping localization of the red and blue channels, and white indicates overlapping localization of all three channels. Bar, 10 μm.