Figure S4.

Additional TGN proteins resemble Drs2 in their kinetic signatures. (A) Maturation kinetics of Drs2 compared with Ste13. A strain expressing HaloTag-Drs2 and the dipeptidyl aminopeptidase GFP-Ste13 was grown to mid-log phase, labeled with JFX dye, and imaged by 4D confocal microscopy. Shown are average projected Z-stacks at the indicated time points from part 5 of Video 4. The upper row shows the complete projections, the second row shows edited projections that include only the cisterna being tracked, and the subsequent rows show the individual fluorescence channels from the edited projections. Scale bar, 2 µm. (B) Quantification of tagged Golgi proteins during a typical maturation event. Depicted are the normalized fluorescence intensities in arbitrary units for the cisterna tracked in A. (C) Smoothed and averaged traces showing the relative kinetic signatures of Drs2 and Ste13. Data were obtained for 10 representative cisternae. (D) Maturation kinetics of Drs2 compared with Stv1. The experiment was performed as in A, except that the strain expressed the proton-pumping ATPase subunit Stv1-GFP, and the images are from part 6 of Video 4. Scale bar, 2 µm. (E) Quantification of tagged Golgi proteins during a typical maturation event. Depicted are the normalized fluorescence intensities in arbitrary units for the cisterna tracked in D. (F) Smoothed and averaged traces showing the relative kinetic signatures of Drs2 and Stv1. Data were obtained for 15 representative cisternae.