The yeast endocytic early/sorting compartment exists as an independent sub-compartment within the trans-Golgi network
- School of Health Science, Tokyo University of Technology, Japan
- Department of Biological Science and Technology, Tokyo University of Science, Japan
- Live Cell Super-Resolution Imaging Research Team, RIKEN Center for Advanced Photonics, Japan
- Institute of Science and Technology Austria, Austria
Figures
Figure 1 with 1 supplement
Localization of endocytosed α-factor at the Tlg2p-residing compartment.
(A) 2D imaging of A594-α-factor and GFP-Tlg2p. Arrowheads indicate examples of overlapping localization. Representative fluorescence intensity profiles along a line (direction from ‘a’ to ‘b’) are indicated in the lower panels. (B) Quantification of GFP-Tlg2p, GFP-Tlg1p, and Sec7-GFP overlapping with A594-α-factor. Data show the mean ± SEM from n ≥ 3 experiments (n > 30 puncta for each experiment). Different letters indicate significant differences at p<0.05 between the indicated times (i.e., no significant difference for a vs. a, significant difference for a vs. b with p<0.05), one-way ANOVA with Tukey’s post hoc test. Error bars indicate the standard SD from n ≥ 3 experiments (n ≥ 30 puncta for each experiment). (C) 3D super-resolution confocal live imaging microscopy (SCLIM) imaging of GFP-Tlg2p and Sec7-mCherry. White dashed lines indicate cell edges. (D) Multi-angle magnified 3D views of the boxed area and the representative fluorescence intensity profiles. Line scan as in (A) shown at right. (E) 3D SCLIM imaging of GFP-Tlg2p, Sec7-iRFP, and pHrode-α-factor; boxed areas shown magnified in (F–H). The images were acquired simultaneously at 5 min after pHrode-α-factor internalization. (F) Multi-angle magnified 3D views of the yellow-boxed area in (E). Line scan as in (A) shown at right. (G) Time series of region in the white-boxed area in (E). Arrows and arrowheads denote the appearance and disappearance of each marker. (H) Higher-magnification views of the red-boxed area in (G). Scale bars, 2.5 μm.
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Figure 1—source data 1
Data for graphs presented in Figure 1B.
- https://cdn.elifesciences.org/articles/84850/elife-84850-fig1-data1-v1.xlsx
Figure 1—figure supplement 1
Localization of α-factor, Sec7p, Tlg1p, and Tlg2p in wild-type cells.
(A) 2D imaging of GFP-Tlg2p and FM4-64 in wild-type cells. (B, C) 2D imaging of A594-α-factor and Sec7-GFP (B) or GFP-Tlg1p (C) in wild-type cells. The images were acquired at 5, 10, or 20 min after A594-α-factor internalization. Yellow arrowheads indicate examples of overlapping localization. Red and green arrowheads indicate example of A594-α-factor or GFP signal, respectively. Higher-magnification views of the boxed area are shown in the right panels. Representative fluorescence intensity profiles along a line (direction from ‘a’ to ‘b’) in the merged images are indicated in the lower panels. (D) 2D imaging of GFP-Tlg1p and mCherry-Tlg2p in wild-type cells shown also in DIC at right. Yellow arrowheads indicate examples of overlapping localization. Red arrowheads indicate example of puncta predominantly localizing mCherry-Tlg2p. Higher-magnification views of the boxed area are shown in the right panels. Representative fluorescence intensity profiles along a line (direction from ‘a’ to ‘b’) in the merged images are indicated in the lower panels. (E) 2D imaging of A647-α-factor, GFP-Tlg1p, and mCherry-Tlg2p. The images were acquired at 5 min after A647-α-factor internalization. Yellow and white arrowheads indicate examples of puncta in which all fluorescent signals overlap. (F) Plates showing the growth phenotype of wild-type cells expressing pRS316 (vector only) or pRS-316-Sec7-iRFP. A dilution series of indicated cells were plated on SD plate lacking uracil and incubated at 25oC and 37oC. Scale bars, 2.5 μm.
Figure 2 with 1 supplement
Dynamic behavior of the Tlg2p-residing compartment, endocytic vesicles, and cargos.
(A, B) 2D imaging of GFP-Tlg2p and Abp1-mCherry in wild-type (A) and arp3-D11A cells (B).Kymographs along lines in the upper merged image are shown in the panels below. A time series of the boxed area in (A) and (B) is shown under the kymograph. Arrowheads highlight the movement of Abp1p toward the Tlg2p sub-compartment. (C–F) 4D super-resolution confocal live imaging microscopy (SCLIM) imaging of GFP-Tlg2p and Abp1-mCherry in wild-type (C, E) and arp3-D11A (D, F). (E, F) Time series of the areas boxed in (C) and (D) are shown in multi-angle magnified 3D views. White and yellow arrowheads indicate the dynamics of different Abp1p patches. (G) 4D SCLIM imaging of GFP-Vps21p and Abp1-mCherry in wild-type cells. Time series of the boxed areas are shown in the right panels. White and red arrowheads indicate the dynamics of the Vps21p-residing endosome and Abp1p patch. (H) The percentages of Abp1p patches that disappeared from the Tlg2p- or Vps21p-residing compartment. Error bars indicate the SD from n ≥ 10 biological replicates (n ≥ 40 Abp1p patches for each experiment). ***p<0.001, unpaired t-test with Welch’s correction. (I) 4D SCLIM imaging of wild-type cells expressing GFP-Tlg2p, Sec7-iRFP, and Abp1-mCherry. (J) Time series of the boxed area in (I) are shown in magnified 3D views. Arrowheads indicate the incorporation of Abp1p patches to the sub-compartment including GFP-Tlg2p and Sec7-iRFP signals. Scale bars, 2.5μm.
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Figure 2—source data 1
Data for graphs presented in Figure 2H.
- https://cdn.elifesciences.org/articles/84850/elife-84850-fig2-data1-v1.xlsx
Figure 2—figure supplement 1
Functionality of Abp1-mCherry and dynamics of Abp1-mCherry patch.
(A) Plates showing the growth phenotype of wild-type cells, sla1Δ and sla1Δ cells expressing Abp1p-mCherry. A dilution series of indicated cells were plated on YPD plate and incubated at 25°C and 37°C. (B) 3D super-resolution confocal live imaging microscopy (SCLIM) imaging of wild-type cells expressing GFP-Tlg2p, Sec7-iRFP, and Abp1-mCherry. (C) Time series of area boxed in red (B) are shown in magnified 3D views. Red and yellow arrowheads indicate Abp1p-mCherry patches fusing with each other. (D) Time series of single patches in area boxed in yellow (B). Scale bars, 2.5 μm.
Figure 3
Alexa-α-factor is transported from the Tlg2p-residing compartment to the Vps21p-residing endosomal compartment.
(A) 2D imaging of GFP-Tlg2p or Sec7-mCherry and mCherry/GFP-Vps21p. (B) Quantification of Tlg2p or Sec7p overlapping with Vps21p. Error bars indicate the SD from n ≥ 3 experiments (n≥30 puncta for each experiment). (C) Time series of the region in the boxed area in (A). Representative fluorescence intensity profiles along a line in the merged image at 8s are shown on the right. Yellow arrowhead indicates overlapping localization. (D) 4D super-resolution confocal live imaging microscopy (SCLIM) imaging of GFP-Tlg2p, Sec7-iRFP, and mCherry-Vps21p. Arrowheads indicate examples of the association of GFP-Tlg2p and mCherry-Vps21p. (E) Magnified views from the time-series in (F). Arrowheads indicate a Vps21p-residing endosome. Representative fluorescence intensity profiles along a line in the merged images at 15s are shown to the right. (F) Time series of the region in the boxed area in (D). (G) Further magnified views from the time series in (F). (H) 2D imaging of A647-α-factor, GFP-Tlg2p, and mCherry-Vps21p. The images were acquired at 5 and 15min after A647-α-factor internalization. Higher-magnification views of the boxed areas are shown in the lower panels. (I) Quantification of A647-α-factor overlapping with GFP-Tlg2p or mCherry-Vps21p. Data show the mean ± SEM from n ≥ 3 experiments (n > 30 puncta for each experiment). Comparisons are made between the same colors, with different letters indicating significant difference (p<0.05) between the indicated times, one-way ANOVA with Tukey’s post hoc test. (J) 2D imaging of A594-α-factor and GFP-Tlg2p in vps21Δ ypt52Δ cells. The images were acquired at 5 and 20min after A594-α-factor internalization. Higher-magnification views of the boxed area are shown in the right panels. Arrowheads indicate examples of the overlapping localization of A594-α-factor and GFP-Tlg2p. (K) Quantification of GFP-Tlg2p overlapping with A594-α-factor. Error bars indicate the SD from n ≥ 3 experiments (n > 30 puncta for each experiment). *p<0.05, unpaired t-test with Welch’s correction. Scale bars, 2.5μm.
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Figure 3—source data 1
Data for graphs presented in Figure 3B.
- https://cdn.elifesciences.org/articles/84850/elife-84850-fig3-data1-v1.xlsx
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Figure 3—source data 2
Data for graphs presented in Figure 3I.
- https://cdn.elifesciences.org/articles/84850/elife-84850-fig3-data2-v1.xlsx
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Figure 3—source data 3
Data for graphs presented in Figure 3K.
- https://cdn.elifesciences.org/articles/84850/elife-84850-fig3-data3-v1.xlsx
Figure 4
GGA adaptors are required for export of A594-α-factor out of the Tlg2p-residing compartment.
(A) 2D imaging of A594-α-factor in cells lacking clathrin adaptor proteins. (B) 2D imaging of A594-α-factor and Sec7-GFP in cells lacking clathrin adaptor proteins. The images were acquired simultaneously at 20min after A594-α-factor internalization. Arrowheads indicate examples of overlapping localization. (C) Quantification of Sec7-GFP overlapping with A594-α-factor. Error bars indicate the SD from n ≥ 3 experiments (n > 30 puncta for each experiment). ***p<0.001, unpaired t-test with Welch’s correction. (D) 2D imaging of A594-α-factor and GFP-Tlg2p in gga1Δ gga2Δ cells. Higher-magnification views of the boxed area are shown in the right panels. Representative fluorescence intensity profiles along lines in the merged images are indicated in the right panels. (E) Quantification of GFP-Tlg2p overlapping with A594-α-factor in gga1Δ gga2Δ cells. The bars surrounded by red lines indicate the total ratio of the Tlg2p sub-compartment overlapping with α-factor. Error bars indicate the SD from n ≥ 3 experiments (n > 30 puncta for each experiment). (F) 3D SCLIM imaging of GFP-Tlg2p and pHrode-α-factor in gga1Δ gga2Δ cells. The images were acquired simultaneously at 20min after pHrode-α-factor internalization. (G, H) Multi-angle magnified 3D views of the boxed areas in (F), representing co-localization (G) or adjacent localization (H) of GFP-Tlg2p and pHrode-α-factor. (I) 3D SCLIM imaging of GFP-Tlg2p, Sec7-iRFP, and pHrode-α-factor in gga1Δ gga2Δ cells. The images were acquired simultaneously at 10min after pHrode-α-factor internalization. (J) Multi-angle magnified 3D views of the boxed area in (I). Scale bars, 2.5μm.
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Figure 4—source data 1
Data for graphs presented in Figure 4C.
- https://cdn.elifesciences.org/articles/84850/elife-84850-fig4-data1-v1.xlsx
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Figure 4—source data 2
Data for graphs presented in Figure 4E.
- https://cdn.elifesciences.org/articles/84850/elife-84850-fig4-data2-v1.xlsx
Figure 5 with 2 supplements
The transition from the Tlg2p- to the Sec7p-residing compartment requires GGA adaptors.
(A) 2D imaging of GFP-Tlg2p and Sec7-mCherry in gga1Δ gga2Δ cells. Representative intensity profiles of GFP-Tlg2p or Sec7-mCherry along a line in the merged images are indicated in the right lower panel. (B) Quantification of GFP-Tlg2p overlapping with Sec7-mCherry in wild-type and gga1Δ gga2Δ cells. Error bars indicate the SD from n ≥ 3 experiments (n > 30 puncta for each experiment). **p<0.01, unpaired t-test with Welch’s correction. (C, E) 4D super-resolution confocal live imaging microscopy (SCLIM) imaging of GFP-Tlg2p and Sec7-mCherry in wild-type cells (C) and gga1Δ gga2Δ cells (E).The time series of regions in the boxed areas in (C) are shown in the lower panels. Arrows and arrowheads denote the appearance and disappearance of each marker. (D, G) Time-course changes in relative fluorescence intensity of GFP-Tlg2p and Sec7-mCherry in the boxed areas in (C) or (E). (F) Time series of the region in the boxed area in (E). (H, I) Multi-angle magnified 3D views from (F). (J) 4D SCLIM imaging of GFP-Tlg2p, Gga2-mCherry, and Sec7-iRFP. The time series of regions in the boxed areas in (J) are shown in the lower panels. (K, L) Multi-angle magnified 3D views of time points from (J).Scale bars, 2.5μm.
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Figure 5—source data 1
Data for graphs presented in Figure 5B.
- https://cdn.elifesciences.org/articles/84850/elife-84850-fig5-data1-v1.xlsx
Figure 5—figure supplement 1
Localization of Tlg2p and Sec7p at different cell cycle stages.
(A) Maximum intensity projections of Z stacks of S- or M-phase cell expressing Sec7-mCherry. The Z series was acquired through the entire cell at 0.2 μm intervals. Cells were synchronized at S- or M-phase by treating with 30 mM hydroxyurea or 50 μM nocodazole for 3 hr at 25°C. (B) Quantification of the number of Sec7-mCherry-positive puncta. Data show the mean ± SD (n = 10 cells). *p<0.05, unpaired t-test with Welch’s correction. (C) 2D imaging of GFP-Tlg2p and Sec7-mCherry in S- or M-phase cell. (D) Quantification of GFP-Tlg2p overlapping with Sec7-mCherry. Data show the mean ± SEM from n ≥ 3 experiments (n > 30 puncta for each experiment). (E) Lifetime of Sec7-mCherry in wild-type and gga1Δ gga2Δ cells. Data show the mean ± SD (n > 10 cisterna for each strain). **p<0.01, unpaired t-test with Welch’s correction.
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Figure 5—figure supplement 1—source data 1
Data for graphs presented in Figure 5—figure supplement 1B.
- https://cdn.elifesciences.org/articles/84850/elife-84850-fig5-figsupp1-data1-v1.xlsx
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Figure 5—figure supplement 1—source data 2
Data for graphs presented in Figure 5—figure supplement 1D.
- https://cdn.elifesciences.org/articles/84850/elife-84850-fig5-figsupp1-data2-v1.xlsx
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Figure 5—figure supplement 1—source data 3
Data for graphs presented in Figure 5—figure supplement 1E.
- https://cdn.elifesciences.org/articles/84850/elife-84850-fig5-figsupp1-data3-v1.xlsx
Figure 5—figure supplement 2
Dynamics of Tlg1p and Tlg2p in wild-type and gga1Δ gga2Δ cells.
(A, D) 2D imaging of GFP-Tlg1p and mCherry-Tlg2p in wild-type (A) or gga1Δ gga2Δ (D) cell. (B, E) A time series of the region boxed in (A, D) are shown. Arrows and arrowheads denote the appearance and disappearance of each marker. (C, F) Time course changes in relative fluorescence intensity of GFP-Tlg1p and mCherry-Tlg2p in wild-type (C) or gga1Δ gga2Δ (F) cell. (G) Plates showing the growth phenotype of gga1Δ cells expressing Gga2-mCherry. A dilution series of indicated cells were plated on YPD plate and incubated at 25°C and 37°C. (H) 2D imaging of GFP-Tlg2p and Gga2-mCherry. Higher-magnification views of the boxed area are shown in the right panels. Representative fluorescence intensity profiles along a line (direction from ‘a’ to ‘b’) in the merged images are indicated in the right panel. Quantification of Gga2-mCherry overlapping with GFP-Tlg2p in wild-type cells is shown in the right panel. Scale bars, 2.5 μm.
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Figure 5—figure supplement 2—source data 1
Data for graphs presented in Figure 5—figure supplement 2H.
- https://cdn.elifesciences.org/articles/84850/elife-84850-fig5-figsupp2-data1-v1.xlsx
Figure 6 with 1 supplement
Snc1p is sorted to the plasma membrane (PM) via the Tlg2p-residing compartment.
(A) Total internal reflection fluorescence (TIRF) imaging of GFP-Snc1p and A594-α-factor in wild-type cells. The images were acquired simultaneously at 5 min after A594-α-factor internalization. Arrowheads indicate A594-α-factor puncta including GFP-Snc1p. (B–E) 4D super-resolution confocal live imaging microscopy (SCLIM) imaging of GFP-Tlg2p, mCherry-Snc1p, and Sec7-iRFP in wild-type cells. (C) Multi-angle magnified 3D views from the 70 s image in (D). Representative fluorescence intensity profiles along lines (direction from ‘a’ to ‘b’) in the merged images are indicated in the right panels. (D) Time series of regions in the boxed area in (B). (E) Higher-magnification views of the indicated time points. (F–H) 4D SCLIM imaging of GFP-Tlg2p, mCherry-Snc1p and Sec7-iRFP in gga1Δ gga2Δ cells. (G) Multi-angle magnified 3D views and representative fluorescence intensity profiles at 55 s in (H). (H) The time series of region in the boxed area in (F). (I) 3D SCLIM imaging of GFP-Tlg2p, mCherry-Snc1p, and Sec7-iRFP in rcy1Δ cells. Multi-angle magnified 3D views of the boxed area and representative fluorescence intensity profiles shown in the lower panels. Scale bars, 2.5 μm.
Figure 6—figure supplement 1
Localization of Snc1p at the Tlg2p-residing compartment.
(A) 2D imaging of GFP-Tlg2p and mCherry-Snc1p in wild-type and gga1Δ gga2Δ cells. Higher-magnification views of the boxed area are shown in the right panels. Representative fluorescence intensity profiles along a line (direction from ‘a’ to ‘b’) in the merged images are indicated in the lower panels. Yellow arrowheads indicate examples of overlapping localization. (B) Quantification of Snc1p overlapping with Tlg2p is shown to the right. Error bars indicate the SD from n ≥ 3 experiments (n > 30 puncta for each experiment). (C) 3D super-resolution confocal live imaging microscopy (SCLIM) imaging of mCherry-Snc1p in wild-type cells. Scale bars, 2.5 μm.
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Figure 6—figure supplement 1—source data 1
Data for graphs presented in Figure 6—figure supplement 1B.
- https://cdn.elifesciences.org/articles/84850/elife-84850-fig6-figsupp1-data1-v1.xlsx
Figure 7 with 1 supplement
Ypt31p is localized at both the Tlg2p-residing compartment and the Sec7p-residing compartment.
(A–D) 4D super-resolution confocal live imaging microscopy (SCLIM) imaging of GFP-Tlg2p, mCherry-Ypt31p, and Sec7-iRFP in wild-type cells. (B) Time series of regions in the boxed area in (A). Arrows and arrowheads denote the appearance and disappearance of each marker. (C) Multi-angle magnified 3D views from the 60 s image in (B). Representative fluorescence intensity profiles along lines (direction from ‘a’ to ‘b’) in the merged images are indicated in the right panels. (D) Higher-magnification views of the red-boxed area in (B). (E–H) 4D SCLIM imaging of GFP-Ypt31p, mCherry-Snc1p and Sec7-iRFP in wild-type cells. (F) Time series of regions in the boxed area in (E). Arrows and arrowheads denote the appearance and disappearance of each marker. (G) Multi-angle magnified 3D views from the 45 s image in (F). Representative fluorescence intensity profiles along lines (direction from ‘a’ to ‘b’) in the merged images are indicated in the right panels. (H) Higher-magnification views of the red-boxed area in (F). (I–L) 4D SCLIM imaging of GFP-Ypt31p, mCherry-Snc1p, and Sec7-iRFP in gga1Δ gga2Δ cells. (J) Time series of regions in the boxed area in (I). Arrows and arrowheads denote the appearance and disappearance of each marker. (K) Multi-angle magnified 3D views from the 55 s image in (J). Representative fluorescence intensity profiles along lines (direction from ‘a’ to ‘b’) in the merged images are indicated in the right panels. (L) Higher-magnification views of the red-boxed area in (J). Scale bars, 2.5 μm.
Figure 7—figure supplement 1
Localization of Ypt31p, Ypt32p, and α-factor in wild-type cell.
(A) Plates showing the growth phenotype of ypt32Δ cells expressing GFP-Ypt31p. A dilution series of indicated cells were plated on YPD plate and incubated at 25°C. (B–D) 4D super-resolution confocal live imaging microscopy (SCLIM) imaging of GFP-Ypt31p and mCherry-Ypt32p in wild-type cell. (C) Time series of regions in the boxed area in (B). (D) Multi-angle magnified 3D views from the 70 s image in (C). Representative fluorescence intensity profiles along lines (direction from ‘a’ to ‘b’) in the merged images are indicated in the right panels. (E) 2D imaging of A594-α-factor and GFP-Ypt31p in wild-type cells. The images were acquired at 5, 10, or 20 min after A594-α-factor internalization. Red and green arrowheads indicate example of A594-α-factor or GFP signal, respectively. Higher-magnification views of the boxed area are shown in the right panels. Representative fluorescence intensity profiles along a line (direction from ‘a’ to ‘b’) in the merged images are indicated in the lower panels. (F) Quantification of GFP-Ypt31p overlapping with A594-α-factor. Data show the mean ± SEM from n ≥ 3 experiments (n > 30 puncta for each experiment).
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Figure 7—figure supplement 1—source data 1
Data for graphs presented in Figure 7—figure supplement 1F.
- https://cdn.elifesciences.org/articles/84850/elife-84850-fig7-figsupp1-data1-v1.xlsx
Figure 8
Model showing the role of the Tlg2p-residing compartment as an early/sorting compartment in the endocytic pathway.
Schematic showing the Tlg2p-residing area as a discrete early/recycling sub-compartment that sorts endocytic cargo to the endocytic or recycling pathway. The effects of gga1Δ gga2Δ cells on the post-TGN trafficking pathway are shown on the right. See details in the text.
Videos
Video 1
Left: multi-angle 3D reconstructed movie of GFP-Tlg2p (green) and Sec7-mCherry (red) in wild-type cell.
Right: multi-angle 3D reconstructed movie of GFP-Tlg2p (green), Sec7-iRFP (red), and pHrodo-α-factor (blue).
Video 2
Triple-color 4D movie of GFP-Tlg2p (green), Sec7-iRFP (red), and pHrodo-α-factor (blue) in a wild-type cell.
Arrows indicate examples of the sequential appearance and disappearance of each protein.
Video 3
Upper video: 2D time-lapse movie of Abp1-mCherry (red in merge) and GFP-Tlg2p (green in merge) in wild-type cell.
Arrows indicate movement of an Abp1-mCherry-labeled endocytic vesicle toward the GFP-Tlg2p-labeled sub-compartment. Lower video: 2D time-lapse movie of Abp1-mCherry (red in merge) and GFP-Tlg2p (green in merge) in the arp3-D11A mutant. Arrows indicate movement of the GFP-Tlg2p-labeled sub-compartment toward an Abp1-mCherry-labeled endocytic vesicle.
Video 4
Dual-color 4D movie of GFP-Tlg2p (green) and Abp1-mCherry (red) in a wild-type cell.
Arrows indicate examples of Abp1-mCherry-labeled vesicles disappearing on the GFP-Tlg2p-labeled sub-compartment.
Video 5
Dual-color 4D movie of GFP-Tlg2p (green) and Abp1-mCherry (red) in the arp3-D11A mutant.
Arrows indicate examples of Abp1-mCherry-labeled vesicles disappearing onto the GFP-Tlg2p-residing sub-compartment.
Video 6
Triple-color 4D movie of GFP-Tlg2p (green), mCherry-Vps21p (red), and Sec7-iRFP (blue) in a wild-type cell.
Arrows indicate examples of association between GFP-Tlg2p and mCherry-Vps21p.
Video 7
Multi-angle 3D reconstructed movie of GFP-Tlg2p (green), Sec7-iRFP (red), and pHrodo-α-factor (blue) in the gga1Δ gga2Δ mutant.
Video 8
Dual-color 4D movie of GFP-Tlg2p (green) and Sec7-mCherry (red) in wild-type cell.
Arrows indicate examples of sequential appearance and disappearance of each protein.
Video 9
Dual-color 4D movie of GFP-Tlg2p (green) and Sec7-mCherry (red) in the gga1Δ gga2Δ mutant.
Video 10
Multi-angle 3D reconstructed movie of GFP-Tlg2p (green) and Sec7-mCherry (red) in the gga1Δ gga2Δ mutant.
Video 11
Triple-color 4D movie of GFP-Tlg2p (green), Sec7-iRFP (red), and Gga2-mCherry (blue) in a wild-type cell.
Arrows indicate examples of the sequential appearance and disappearance of each protein.
Video 12
Multi-angle 3D reconstructed movie of GFP-Tlg2p (green), Sec7-iRFP (red), and mCherry-Snc1p (blue) in the gga1Δ gga2Δ mutant.
Video 13
Multi-angle 3D reconstructed movie of GFP-Tlg2p (green), Sec7-iRFP (red), and mCherry-Snc1p (blue) in the rcy1Δ mutant.
Additional files
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Supplementary file 1
List of strains used in this study.
- https://cdn.elifesciences.org/articles/84850/elife-84850-supp1-v1.doc
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MDAR checklist
- https://cdn.elifesciences.org/articles/84850/elife-84850-mdarchecklist1-v1.pdf
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The yeast endocytic early/sorting compartment exists as an independent sub-compartment within the trans-Golgi network
eLife 12:e84850.
https://doi.org/10.7554/eLife.84850
