Fig. 5.
The intracellular pool of GFP-nSMase2 is not exclusively associated with the Golgi apparatus. MCF-7 cells were plated at low density (0.3 × 106/dish) and, 24 h later, were transfected with 3′-GFP-tagged nSMase2 (0.5 µg/dish). After 18–20 h, cells were treated with (CHX) or without (none) 50 µg/ml CHX for 2 h and co-localization of 3′-GFP-nSMase2 with giantin was analyzed by confocal microscopy. (A) Co-localization of GFP-nSMase2 with giantin and localization at cell surface was quantified. Results are represented as the percentage of cells displaying localization in the Golgi (Golgi), at the plasma membrane (PM), or showing both localization at the PM and co-localization with giantin (PM and Giantin+) or not (PM and Giantin−). For each condition, at least 100 cells were quantified under the microscope. Values are means±s.e.m. of three independent experiments. (B) Pictures are representative of at least 4 fields taken from four independent experiments.