dNGLUC secretion is Monensin and Brefeldin A-sensitive. (a, b) Monensin and Brefeldin A inhibit non-conventional secretion of dNGLUC (a) and secreted alkaline phosphatase (SEAP) (b). Twenty-four hours after transfection of 293ET with dNGLUC, medium was replaced with medium containing 7 μM Monensin (Mon), 10 μg/ml Brefeldin A (Bref) and 5 μg/ml MG132 or Methanol (Me). SN was collected after 4 h prior to analysis of GLUC or SEAP activity in the supernatant. NT, not transfected; RLU, relative light units.) (c) Co-localization of GFP-dNGLUC and Golgin-67. 293ET cells were transfected with GFP-dNGLUC and Golgin67-DsRed and cells were fixed with 4% paraformaldehyde prior to analysis by confocal microscopy. Overlap of GFP-dNGLUC and Golgin67-DsRed is marked by an arrow.
Ketteler et al. Genome Biology 2008 9:R64 doi:10.1186/gb-2008-9-4-r64