bioimagesUnknownLidia HurtadoThe Journal of Cell Biologyhttps://www.ebi.ac.uk/biostudies/studies/S-JCBD-201011014JCB

Reassembly of the GA after NZ washout. RPE1 cells co-expressing GT-m-cherry and GFP-AK1B were treated with NZ, and labeling for GMAP210 (green) and gamma-tubulin (red) is shown. Time after NZ washout: 180 min.

RPE-1 cells were cotransfected with VSV-GFP and flag-AK1. After transfection, cells were incubated at 40ºC and were then transferred to 32ºC and fixed at 100 min. Cells were labelled for GFP (green) and AKAP450(red).

RPE-1 cells were transfected with VSV-GFP(control). After transfection, cells were incubated at 40ºC and were then transferred to 32ºC and fixed at 60 min. Cells were labelled for GFP (green) and AKAP450(red).

High magnification image of a flag-AK1-transfected cell triple labelled for α-tubulin (green), flag (red) and GMAP210 (blue) to visualize MT network.

A flag-AK1-transfected cell incubated on ice for 40 min to depolymerize MTs was triple stained for α-tubulin (green), flag (red) and GMAP210 (blue).

RPE-1 cells were transfected with VSV-GFP. After transfection, cells were incubated at 40ºC and were then transferred to 32ºC and fixed at 0 min. Cells were labelled for GFP (green) and AKAP450(red).

Subcellular localization of GFP-AK1 (green). A GFP-AK1-expressing cell was labelled for GMAP210 (red).

Low magnification image of a flag-AK1B transfected cells triple labelled for flag (green), AKAP450 (red) and GMAP210 (blue).

Merged image of a flag-AK1-transfected cell stained with α-tubulin (green), flag (red) and GMAP210 (blue) to visualize the MT network.

Control cells were transfected with the flag-AK1B construct and then NZ treated to fragment the GA into stacks. Labellings were for flag (green), GM130 (red) and GMAP210 (blue).

RPE-1 cells were transfected with VSV-GFP(control). After transfection, cells were incubated at 40ºC and were then transferred to 32ºC and fixed at 100 min. Cells were labelled for GFP (green) and AKAP450(red).

RPE-1 cells were cotransfected with VSV-GFP and flag-AK1. After transfection, cells were incubated at 40ºC and were then transferred to 32ºC and fixed at 0 min. Cells were labelled for GFP (green) and AKAP450(red).

GM130-depleted cells were transfected with the flag-AK1B construct and then NZ treated to fragment the GA into stacks. Labelings were for flag (green), GM130 (red) and GMAP210 (blue).

FRAP experiments were performed in RPE-1 GT-GFP expressing cells co-transfected with mcherry-centrin and flag-AK1B. An image of a selected cell is shown before bleaching (pre-bleach) with CTR in red and the GA in green.

RPE-1 cells were cotransfected with VSV-GFP and flag-AK1B. After transfection, cells were incubated at 40ºC and were then transferred to 32ºC and fixed at 0 min. Cells were labelled for GFP (green) and AKAP450(red).

Merged image of a cell expressing flag-AK1B triple labelled for flag (green), GM130 (red) and Golgin245 (blue).

FRAP experiments were performed in RPE-1 GT-GFP expressing cells co-transfected with mcherry-centrin and flag-AK1B. A video from of a selected cell is shown 0 sec after bleaching with CTR in red and the GA in green.

Representative images of control cells maintained in 0.25% serum for 36h and triple labelled for polyglutamylated tubulin (green), AKAP450 (red) and GMAP210 (blue).

Merged image of a flag-AK1B-expressing cell triple labelled for flag (green), AKAP450 (red) and GMAP210 (blue).

Translocation of Venus-NPY containing vesicules in AK1-expressing cells. Images recorded over 2 min were overlaid to visualize the vesicle tracks.

Reorientation of the CTR and the GA after wounding in AK1-transfected cells. Cells were stained with flag (green), AKAP450 (red)and GMAP210 (blue).

Merged image of a cell expressing flag-AK1 triple labelled for flag (green), GM130 (red) and Golgin245 (blue).

FRAP experiments were performed in RPE-1 GT-GFP expressing cells co-transfected with mcherry-centrin and either flag-AK1. An image of a selected cell is shown before bleaching (pre-bleach) with CTR in red and the GA in green.

Merged image of a flag-AK1B-expresssing cell stained with α-tubulin (green), flag (red) and GMAP210 (blue) to visualize the MT network.

A control cell incubated on ice for 40 min to depolymerize MTs and rewarmed for 2 min was triple stained for α-tubulin (green), flag (red) and GMAP210 (blue).

RPE-1 cells were transfected with VSV-GFP (control). VSV-GFP was detected at the PM by incubating with the ectodomain antibody 8G5 during the last 30 min of a 60 or 100 min period after the shift at a permissive temperature. Cells were then fixed with methanol and triple labelled for 8G5 (green), AKAP450 (red) and GMAP210 (blue).

RPE-1 cells were cotransfected with VSV-GFP and flag-AK1B. After transfection, cells were incubated at 40ºC and were then transferred to 32ºC and fixed at 60 min. Cells were labelled for GFP (green) and AKAP450(red).

RPE-1 cells were cotransfected with VSV-GFP and flag-AK1. VSV-GFP was detected at the PM by incubating with the ectodomain antibody 8G5 during the last 30 min o a 60 or 100 min period after the shift at a permissive temperature. Cells were then fixed and triple labelled for 8G5 (green), AKAP450(red) and GMAP210 (blue).

FRAP experiments were performed in RPE-1 GT-GFP expressing cells co-transfected with mcherry-centrin and flag-AK1B. A video frame of a selected cell is shown 200 sec after bleaching with CTR in red and the GA in green.

A flag-AK1-transfected cell incubated on ice for 40 min to depolymerize MTs and rewarmed for 7 min was triple labelled for α-tubulin (green), flag (red) and GMAP210 (blue).

Control cells were treated with NZ to depolymerise MTs. At 15 min after washout, cells were triple labelled for α-tubulin (green), flag (red) and GMAP210 (blue).

Reassembly of the GA after NZ washout. RPE1 cells co-expressing GT-m-cherry and GFP-AK1 were treated with NZ, and labeling for GMAP210 (green) and gamma-tubulin (red) is shown. Time after NZ washout: 0 min.

Flag-AK1B transfected cells were treated with NZ to depolymerise MTs. Before washout, cells were triple labelled for α-tubulin (green), flag (red) and GMAP210 (blue).

Reorientation of the CTR and the GA after wounding in AK1B-transfected cells. Cells were stained with flag (green), AKAP450 (red) and GMAP210 (blue).

A control cell incubated on ice for 40 min to depolymerize MTs and rewarmed for 7 min was triple stained for α-tubulin (green), flag (red) and GMAP210 (blue).

Representative images flag-AK1 transfected cells maintained in 0.25% serum for 36h and triple labelled for polyglutamylated tubulin (green), AKAP450 (red) and GMAP210 (blue).

FRAP experiments were performed in RPE-1 GT-GFP expressing cells co-transfected with mcherry-centrin and flag-AK1. A video frame of a selected cell is shown 200 sec after bleaching with CTR in red and the GA in green.

Reassembly of the GA after NZ washout. RPE1 cells expressing GT-mcherry were treated with NZ, and labeling for GMAP210 (green) and gamma-tubulin (red) is shown. Time after NZ washout: 180 min.

A flag-AK1-transfected cell incubated on ice for 40 min to depolymerize MTs and rewarmed for 2 min was triple stained for α-tubulin (green), flag (red) and GMAP210 (blue).

Flag-AK1B transfected cells were treated with NZ to depolymerise MTs. At 5 min after washout, cells were triple labelled for α-tubulin (green), flag (red) and GMAP210 (blue).

FRAP experiments were performed in RPE-1 GT-GFP expressing cells transfected with mcherry-centrin. A video frame of a selected cells is shown 200 sec after bleaching with CTR in red and the GA in green.

Representative images flag-AK1B transfected cells maintained in 0.25% serum for 36h and triple labelled for polyglutamylated tubulin (green), AKAP450 (red) and GMAP210 (blue).

Low magnification image of a flag-AK1 transfected cells triple labelled for flag (green), AKAP450 (red) and GMAP210 (blue).

FRAP experiments were performed in RPE-1 GT-GFP expressing cells co-transfected with mcherry-centrin and flag-AK1. A video frame of a selected cell is shown 0 sec after bleaching with CTR in red and the GA in green.

FRAP experiments were performed in RPE-1 GT-GFP expressing cells transfected with mcherry-centrin. An image of a selected cell is shown before bleaching (pre-bleach) with CTR in red and the GA in green.

RPE-1 cells were cotransfected with VSV-GFP and flag-AK1B. After transfection, cells were incubated at 40ºC and were then transferred to 32ºC and fixed at 100 min. Cells were labelled for GFP (green) and AKAP450(red).

RPE-1 cell expressing the flag-AK1 fragment was triple labelled for flag (green), AKAP450 (red) and GMAP210 (blue).

Control cells were treated with NZ to depolymerise MTs. At 5 min after washout, cells were triple labelled for α-tubulin (green), flag (red) and GMAP210 (blue).

Reassembly of the GA after NZ washout. RPE1 cells co-expressing GT-m-cherry and GFP-AK1B were treated with NZ, and labeling for GMAP210 (green) and gamma-tubulin (red) is shown. Time after NZ washout: 0 min.

RPE-1 cells were cotransfected with VSV-GFP and flag-AK1. After transfection, cells were incubated at 40ºC and were then transferred to 32ºC and fixed at 60 min. Cells were labelled for GFP (green) and AKAP450(red).

A control cell incubated on ice for 40 min to depolymerize MTs was triple stained for α-tubulin (green), flag (red) and GMAP210 (blue).

RPE-1 cells were cotransfected with VSV-GFP and flag-AK1B. VSV-GFP was detected at the PM by incubating with the ectodomain antibody 8G5 during the last 30 min o a 60 or 100 min period after the shift at a permissive temperature. Cells were then fixed and triple labelled for 8G5 (green), AKAP450 (red) and GMAP210 (blue).

Reassembly of the GA after NZ washout. RPE1 cells co-expressing GT-m-cherry and GFP-AK1 were treated with NZ, and labeling for GMAP210 (green) and gamma-tubulin (red) is shown. Time after NZ washout: 180 min.

FRAP experiments were performed in RPE-1 GT-GFP expressing cells transfected with mcherry-centrin. A video frame of a selected cell is shown 0 sec after bleaching (pre-bleach) with CTR in red and the GA in gree.

Translocation of Venus-NPY containing vesicules in AK1B-expressing cells. Images recorded over 2 min were overlaid to visualize the vesicle tracks.

Control cells were treated with NZ to depolymerise MTs. Before washout, cells were triple labelled for α-tubulin (green), flag (red) and GMAP210 (blue).

Reassembly of the GA after NZ washout. RPE1 cells expressing GT-mcherry were treated with NZ, and labeling for GMAP210 (green) and gamma-tubulin (red) is shown. Time after NZ washout: 0 min.

Subcellular localization of GFP-AK4 (green). A GFP-AK4 expressing cell was labelled for γ-tubulin (red).

Flag-AK1B transfected cells were treated with NZ to depolymerise MTs. At 15 min after washout, cells were triple labelled for α-tubulin (green), flag (red) and GMAP210 (blue).

Translocation of Venus-NPY containing vesicules in control cells. Images recorded over 2 min were overlaid to visualize the vesicle tracks.

Subcellular localization of GFP-AK1B (green). A GFP-AK1B expressing cell was labelled for GMAP210 (red).

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