Fig. 5: NSC80997 and NSC255112 trigger reversible Golgi fragmentation.

HeLa^WT cells were treated with 10 µM NSC80997, NSC255112 or Brefeldin A and processed for immunofluorescence. a–c HeLa cells were imaged by confocal microscopy to reveal the morphology of the Golgi apparatus (giantin, green) and the nuclei (DAPI, magenta). a Time course of Golgi fragmentation during treatment with NSC80997. Scale bars represent 10 µm. b, c After treatment for 24 h with either NSC80997 (b) or NSC255112 (c), cells were washed with fresh medium and incubated for another 24 hrs without inhibitor. Scale bar represents 20 µm. d Bar plot shows percentage of HeLa^WT cells displaying compact, fragmented or diffuse Golgi morphologies at different timepoints during treatment with NSC80997. Data is presented as mean + SEM from three independent experiments with 62–109 cells/experiment. Example images depicting compact, fragmented or diffuse Golgi morphologies are shown. Scale bars represent 10 µm. e Line graph depicts Pearson’s correlation coefficient of bCOP (magenta) versus cis-Golgi marker GM130 (green) at different timepoints during treatment with NSC80997 or Brefeldin A. Images were acquired as Z-stacks using spinning-disk microscopy and data is presented as mean ± SEM from three independent experiments with 10-15 cells/experiment. Representative images of co-localization are shown. Scale bars represent 10 µm.