Pharmacological inhibition of SMS causes extensive tubulation of cis Golgi and TGN in INS-1 cells.
INS-1 cells were treated with DMSO and water (control), and 20 µM of PKD inhibitor or 30 µg/ml of SMS inhibitor D609. After 4 hours of incubation, cells were fixed and processed for indirect Immunofluorescence with rat anti TGN 38 (TGN marker) mouse monoclonal antibodies and anti mouse Alexa Fluor®555-conjugated secondary antibodies (red), and anti GPP130 (cis Golgi marker) rabbit polyclonal antibodies and anti rabbit Alexa Fluor® 488-conjugated secondary antibodies (green). Confocal images were captured and processed using LSM 510 META. Colocalization in 3D images was achieved using velocity 3 software (improvision). Images are representatives of at least two independent experiments.