Localization of ATB0,+ and SEC24C in MCF7 cells. Cells were fixed with methanol, as described in Materials and methods. They were incubated, as indicated, with either anti-SEC24C or anti-SEC24D antibodies, detected with Alexa Fluor® 488-conjugated AffinityPure Fab Fragment Goat Anti-Rabbit IgG (H + L) (green). Further treatment was performed as described in Materials and methods Section 2.5. Next the cells were incubated with anti-ATB0,+ antibodies conjugated with biotin, followed by incubation with AlexaFluor568 conjugated streptavidin (red). Nuclei are visualized with DAPI. NC, negative control without the primary antibodies. B0,+, analysis of B0,+ localization without anti-SEC24 antibodies. Bar 20 μm.