Figure S1.
Giantin distribution among ministacks, morphometric parameters of a side-view ministack, and the effect of overexpression on the LQ. (A) HeLa cells expressing GalT-mCherry were nocodazole-treated for 3 h and immunostained for giantin and βCOP. The Airyscan image demonstrates that almost all ministacks are positive for giantin. Box with a solid line, en face view. Box with a dotted line, side-view. Scale bar, 10 µm. (B and C) Diagrams illustrating the morphometric parameters of a side-view ministack. (B) The rim-localized Golgi protein such as giantin. (C) The interior-localized Golgi protein such as GM130. The side-view of the ministack is positioned such that the trans side is up and the Golgi axis is vertical. An imaginary Golgi ministack is schematically overlaid (pink). The axial line intensity profile is acquired along the axial line (green vertical box). The axial position is defined as the fluorescence center or the Gaussian center of the axial line intensity profile. The lateral line intensity profile is acquired along the lateral line (brown horizontal box). The lateral line intensity profile of the rim-localized Golgi protein such as giantin (B) is different from that of the interior-localized one such as GM130 (C). Typically, the former has the two-peak Gaussian distribution, while the latter has the super-Gaussian distribution. The lateral width of a punctum for giantin and the lateral size are schematically illustrated in the diagram. (D) Investigating the effect of overexpression on the LQ. 10 GFP-tagged Golgi proteins were selected for investigation. For each ministack, the LQ was plotted against the corresponding relative expression level of the protein as a red dot. The relative expression level was calculated as the integrated intensity of the protein divided by that of endogenous GM130 in a ministack. It was further normalized such that the lowest value is 1. Black lines were used to connect red dots from low to high relative expression levels. The linear regression fitting line (green) and its formula, adjusted R2 (adj. R2), and n (the number of ministacks analyzed) are shown.

Giantin distribution among ministacks, morphometric parameters of a side-view ministack, and the effect of overexpression on the LQ. (A) HeLa cells expressing GalT-mCherry were nocodazole-treated for 3 h and immunostained for giantin and βCOP. The Airyscan image demonstrates that almost all ministacks are positive for giantin. Box with a solid line, en face view. Box with a dotted line, side-view. Scale bar, 10 µm. (B and C) Diagrams illustrating the morphometric parameters of a side-view ministack. (B) The rim-localized Golgi protein such as giantin. (C) The interior-localized Golgi protein such as GM130. The side-view of the ministack is positioned such that the trans side is up and the Golgi axis is vertical. An imaginary Golgi ministack is schematically overlaid (pink). The axial line intensity profile is acquired along the axial line (green vertical box). The axial position is defined as the fluorescence center or the Gaussian center of the axial line intensity profile. The lateral line intensity profile is acquired along the lateral line (brown horizontal box). The lateral line intensity profile of the rim-localized Golgi protein such as giantin (B) is different from that of the interior-localized one such as GM130 (C). Typically, the former has the two-peak Gaussian distribution, while the latter has the super-Gaussian distribution. The lateral width of a punctum for giantin and the lateral size are schematically illustrated in the diagram. (D) Investigating the effect of overexpression on the LQ. 10 GFP-tagged Golgi proteins were selected for investigation. For each ministack, the LQ was plotted against the corresponding relative expression level of the protein as a red dot. The relative expression level was calculated as the integrated intensity of the protein divided by that of endogenous GM130 in a ministack. It was further normalized such that the lowest value is 1. Black lines were used to connect red dots from low to high relative expression levels. The linear regression fitting line (green) and its formula, adjusted R2 (adj. R2), and n (the number of ministacks analyzed) are shown.

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