Figure 8.
Other transmembrane Golgi proteins resemble Sys1 in their kinetic signatures.(A) Mislocalization of the transmembrane Golgi proteins Aur1 and Rbd2 in apl4Δ ent5Δ cells after CK-666 treatment. The experiment was performed as in Fig. 3 D. Scale bar, 2 µm. (B) Quantification of the effects in A. The experiment was performed as in Fig. 3 E. (C) Maturation kinetics of Drs2 compared with Aur1. A strain expressing HaloTag-Drs2 and Aur1-GFP 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 1 of Video 8. 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. (D) Quantification of tagged Golgi proteins during typical maturation events. Depicted are the normalized fluorescence intensities in arbitrary units for the cisterna tracked in C. (E) Smoothed and averaged traces showing the relative kinetic signatures of Drs2 and Aur1. Data were obtained for 10 representative cisternae. (F) Maturation kinetics of Drs2 compared with Rbd2. The experiment was performed as in C, except with a strain expressing HaloTag-Drs2 and Rbd2-GFP. Shown are average projected Z-stacks at the indicated time points from part 2 of Video 8. Scale bar, 2 µm. (G) Quantification of tagged Golgi proteins during typical maturation events. Depicted are the normalized fluorescence intensities in arbitrary units for the cisterna tracked in F. (H) Smoothed and averaged traces showing the relative kinetic signatures of Drs2 and Rbd2. Data were obtained for 14 representative cisternae. Error bars represent SEM.

Other transmembrane Golgi proteins resemble Sys1 in their kinetic signatures. (A) Mislocalization of the transmembrane Golgi proteins Aur1 and Rbd2 in apl4Δ ent5Δ cells after CK-666 treatment. The experiment was performed as in Fig. 3 D. Scale bar, 2 µm. (B) Quantification of the effects in A. The experiment was performed as in Fig. 3 E. (C) Maturation kinetics of Drs2 compared with Aur1. A strain expressing HaloTag-Drs2 and Aur1-GFP 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 1 of Video 8. 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. (D) Quantification of tagged Golgi proteins during typical maturation events. Depicted are the normalized fluorescence intensities in arbitrary units for the cisterna tracked in C. (E) Smoothed and averaged traces showing the relative kinetic signatures of Drs2 and Aur1. Data were obtained for 10 representative cisternae. (F) Maturation kinetics of Drs2 compared with Rbd2. The experiment was performed as in C, except with a strain expressing HaloTag-Drs2 and Rbd2-GFP. Shown are average projected Z-stacks at the indicated time points from part 2 of Video 8. Scale bar, 2 µm. (G) Quantification of tagged Golgi proteins during typical maturation events. Depicted are the normalized fluorescence intensities in arbitrary units for the cisterna tracked in F. (H) Smoothed and averaged traces showing the relative kinetic signatures of Drs2 and Rbd2. Data were obtained for 14 representative cisternae. Error bars represent SEM.

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