Continuous presence of the fluorescent secretory cargo within maturing Golgi cisternae. Cells expressed the APVNTT-DsRed-Express2-FKBPRD(C22V) cargo, together with GFP-Vrg4 to mark the early Golgi and Sec7-HaloTag to mark the late Golgi. A culture was grown to mid-log phase, labeled with JF646, washed, and mounted in a flow chamber. Medium that contained SLF was added while imaging on a confocal microscope, with a Z-stack collected every 3 s. The top panel shows the complete video, with projections of the full Z-stacks. The second panel shows an edited video of two representative cisternae. The bottom three panels show the individual fluorescence channels. Bright-field images of the cells are present in all panels. Scale bar, 2 µm. Frames from this video are shown in Fig. 6 A.
