-tagRFP → Clc1-GFP Sec7-tagRFP → Apl2-GFP Sec7-tagRFP → Gga2-GFP Clc1-mCherry → Apl2-GFP Gga2-GFP → Clc1-mCherry Sec7-tagRFP → Chs5-GFP Chs5-GFP → Clc1-mCherry GFP-Tlg2 → Sec7-tagRFP GFP-Tlg2 → Clc1-mCherry Sec21-GFP → Sec7-mCherry Sec21-2xmCherry → GFP-Tlg2 Gnt1-GFP → Sec21-2xmCherry Sys1-GFP → Sec7-mCherry Sys1-iRFP → GFP-Tlg2 Sec21-2xmCherry → Sys1-GFP Mnn9-mCherry → Gnt1-GFP Mnn9-mCherry → Sys1-GFP
Table S1. Peak-to-peak duration time. 
‡Number of puncta (n) in total cells (N) Peak-to-peak duration (s) (mean ± SD) *12 47.9 ± 17.7 GFP-Ypt6 → Sec7-mRFP 13 GFP-Ypt6 → Sys1-iRFP ‡Number of puncta and cells used for the calculation of peak-to-peak duration. In each cell, 1 -3 puncta were selected for the calculation. 

Figure S1 .

 S1 Figure S1. 3D co-localization analyses of Golgi/TGN-resident proteins.(A-H) A variety of GFP-tagged Golgi/TGN-resident proteins (Clc1, Gga2, Apl2, Apl6, 


Figure S4 .Fig. 4 ,

 S44 Figure S2. 4D dynamics of Clc1, Apl2, and Gga2 versus Sec7. 


Figure S6 .

 S6 Figure S6. 4D dynamics of Sys1 versus Sec7, Clc1, and Tlg2. 


Figure

  Figure S7. 4D dynamics of Sys1 versus Ypt6, Sec21 versus Sec7, and Sec21 versus 




  Y., Kurokawa, K., Hirata, R., and Nakano, A. 2013. Rab GAP cascade regulates dynamics of Ypt6 in the Golgi traffic. Proc. Natl. Acad. Sci. U S A. 110:18976-18981. †Ishii, M., Suda, Y., Kurokawa, K., and Nakano, A. 2016. COPI is essential for Golgi cisternal maturation and dynamics. J. Cell Sci.129:3251-3261. 




  Fig. 1A-C, S1A, S2A YSY1 with pRS316-ADH1p-SEC7-tagRFP, 43-C1 with pRS316-ADH1p-SEC7-tagRFP Fig. 1D-F, S1C, S2B TTY5 with pRS316-ADH1p-SEC7-tagRFP Fig. 1G-I, S1B, S2C 11-D11 with pRS316-ADH1p-SEC7-tagRFP Fig. 1J-L, S3A TTY11 Fig. 1M-O, S3B TTY27 Fig. 2A,B, S1D, S4A TTY6 with pRS316-ADH1p-SEC7-tagRFP Fig. 2C,D, S4B TTY3, TTY20 Fig. 3A-C, S1E, S4C TTY10 with pRS316-ADH1p-SEC7-tagRFP Fig. 3D-F, S4D TTY21 Fig. 3G-I TTY21 with pRS316-ADH1p-SEC7-iRFP Fig. 4A-C, S1F, S5A 


Table S2 .

 S2 Yeast strains used in this study. Sikorski, R.S., and Hieter, P. 1989. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics. 122:19-27. 2. Brachmann, C.B., Davies, A., Cost, G.J., Caputo, E., Li, J., Hieter, P., and Boeke, J.D. 1998. Designer deletion strains derived from Saccharomyces cerevisiae S288C: a useful set of strains and plasmids for PCR-mediated gene disruption and other applications. 


	Strain	Genotype	Source
	YPH499 MAT a ura3-52 lys2-801 ade2-101 trp1-Δ63 his3-Δ200 leu2-Δ1	1
	BY4741 MAT a his3Δ1 leu2Δ0 met15Δ0 ura3Δ0	2
	43-C1	BY4741 CLC1-GFP(S65T)::HIS3MX6	3
	11-D11 BY4741 GGA2-GFP(S65T)::HIS3MX6	3
	38-E4	BY4741 APL6-GFP(S65T)::HIS3MX6	3
	YSY1	YPH499 ADE2::pRS402 CLC1-GFP(S65T)::HIS3MX6	This study
	YSY112 YPH499 ADE2::pRS402 SEC7-GFP(S65T)::kanMX6	This study
	YSY115 YPH499 ADE2::pRS402 SEC7-mCherry::natNT2	This study
	TTY3	BY4741 APL6-GFP(S65T)::HIS3MX6 CLC1-mCherry::natNT2	This study
	TTY4	YPH499 ADE2::pRS402 CLC1-mCherry::natNT2	This study
	TTY5	YPH499 ADE2::pRS402 APL2-GFP(S65T)::HIS3MX6	This study
	TTY6	YPH499 ADE2::pRS402 APL6-GFP(S65T)::HIS3MX6	This study
	TTY10 YPH499 ADE2::pRS402 CHS5-GFP(S65T)::HIS3MX6	This study
	TTY11 YPH499 ADE2::pRS402 CLC1-mCherry::natNT2 APL2-GFP(S65T)::HIS3MX6	This study
	TTY20 YPH499 ADE2::pRS402 CLC1-mCherry::natNT2 APL6-GFP(S65T)::HIS3MX6	This study
	TTY21 YPH499 ADE2::pRS402 CLC1-mCherry::natNT2 CHS5-GFP(S65T)::HIS3MX6 This study
	TTY24 YPH499 ADE2::pRS402 CLC1-mCherry::natNT2 SEC21-GFP(S65T)::HIS3MX6 This study
	TTY27 YPH499 ADE2::pRS402 CLC1-mCherry::natNT2 GGA2-GFP(S65T)::HIS3MX6 This study
	TTY29 YPH499 ADE2::pRS402 SEC21-2xmCherry::hphNT1	This study
	1. Yeast. 14:115-132.	
	3. Huh, W.K., Falvo, J.V., Gerke, L.C., Carroll, A.S., Howson, R.W., Weissman, J.S., and O'Shea, E.K.
	2003. Global analysis of protein localization in budding yeast. Nature. 425:686-691.	

Table S3 .

 S3 Plasmids used in this study. 


	Plasmid name	Description	Source
			1
			2
			3
			4
			This study
			This study
			This study
			This study
			5

Table S4 .

 S4 Strains and plasmids used in each experiment. 



J. Cell Sci.: doi:10.1242/jcs.231159: Supplementary information 

J. Cell Sci.: doi:10.1242/jcs.231159: Supplementary information Journal of Cell Science • Supplementary information 

J. Cell Sci.: doi:10.1242/jcs.231159: Supplementary information 

Journal of Cell Science • Supplementary information