1. ArfGAP1 expression alters GBF1 recruitment to Golgi membranes.
ArfGAP1 expression alters GBF1 recruitment to Golgi membranes. (A) HeLa cells were mock transfected or transfected with plasmids encoding EGFP-tagged forms of ArfGAP1 WT (GAP1WT), ArfGAP1 RQ (GAP1RQ), ArfGAP2 WT (GAP2WT), ArfGAP2 RQ (GAP2RQ), ArfGAP3 WT (GAP3WT) or ArfGAP3 RQ (GAP3RQ). Eighteen hours post transfection, cells were fixed and stained with mouse anti-GBF1 antibody and imaged using spinning-disc confocal microscopy. Expression levels of all EGFP-tagged proteins appeared similar as judged by signal intensity. Representative images are shown as projections of all z-slices. (B) Quantification was performed as described in Materials and Methods using a minimum of ten cells from each condition from each of the three independent experiments. The graph reports the fraction of endogenous GBF1 localized to the Golgi in cells that were mock-transfected or transfected with the indicated ArfGAPs. Error bars are ±s.d. (n=3). * indicate nuclei. Values were (0.09, 0.13, 0.23) for mock; (0.28, 0.24, 0.322) for GAP1WT; (0.070, 0.039, 0.118) for GAP1RQ; (0.19, 0.19, 0.30) for GAP2WT; (0.13, 0.16, 0.31) for GAP2RQ; (0.15, 0.12, 0.24) for GAP3WT; (0.14, 0.17, 0.30) for GAP3RQ. Scale bar: 26 μm.
Douglas Quilty et al. J Cell Sci 2019;132:jcs208199
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