Figure 5.
Other transmembrane TGN proteins resemble Drs2 in their kinetic signatures.(A) Maturation kinetics of Drs2 compared with Tlg1. A strain expressing GFP-Drs2 and the SNARE HaloTag-Tlg1 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 3 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 Tlg1. Data were obtained for 10 representative cisternae. (D) Maturation kinetics of Drs2 compared with Kex2. The experiment was performed as in A, except with a strain expressing HaloTag-Drs2 and Kex2-GFP. Shown are average projected Z-stacks at the indicated time points from part 4 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 Kex2. Data were obtained for 13 representative cisternae. (G) Maturation kinetics of Drs2 compared with Vrg4. The experiment was performed as in A, except with a strain expressing HaloTag-Drs2 and GFP-Vrg4. Shown are average projected Z-stacks at the indicated time points from part 1 of Video 5. Scale bar, 2 µm. (H) Quantification of tagged Golgi proteins during a typical maturation event. Depicted are the normalized fluorescence intensities in arbitrary units for the cisterna tracked in G. (I) Smoothed and averaged traces showing the relative kinetic signatures of Drs2 and Vrg4. Data were obtained for 12 representative cisternae.

Other transmembrane TGN proteins resemble Drs2 in their kinetic signatures. (A) Maturation kinetics of Drs2 compared with Tlg1. A strain expressing GFP-Drs2 and the SNARE HaloTag-Tlg1 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 3 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 Tlg1. Data were obtained for 10 representative cisternae. (D) Maturation kinetics of Drs2 compared with Kex2. The experiment was performed as in A, except with a strain expressing HaloTag-Drs2 and Kex2-GFP. Shown are average projected Z-stacks at the indicated time points from part 4 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 Kex2. Data were obtained for 13 representative cisternae. (G) Maturation kinetics of Drs2 compared with Vrg4. The experiment was performed as in A, except with a strain expressing HaloTag-Drs2 and GFP-Vrg4. Shown are average projected Z-stacks at the indicated time points from part 1 of Video 5. Scale bar, 2 µm. (H) Quantification of tagged Golgi proteins during a typical maturation event. Depicted are the normalized fluorescence intensities in arbitrary units for the cisterna tracked in G. (I) Smoothed and averaged traces showing the relative kinetic signatures of Drs2 and Vrg4. Data were obtained for 12 representative cisternae.

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