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Files in this Data Supplement:
Fig. S1. Calculation of the polarization factor. (A,F) The transmission or fluorescent image of the cell is acquired, and threshold and its cell contour is extracted. (B) All cell contours are centred according to the centre of mass and overlay or aligned along their major axis prior to centering (G). Cell contours are fitted using an ellipse (C,D and H,I). The cell contours are added up and the mean size of the length and the width is measured (E and J). An identical approach is used to measure the nuclear polarization factor using the nucleus as an entry point of the analysis process. All steps have been done using a Macro in ImageJ downloadable at http://rsb.info.nih.gov/ij/
Fig. S2. Calculation of the Golgi complex and the nucleus distance. (A) The fluorescent image of the nucleus, Golgi complex and cell membrane are acquired and threshold. (B) Images are segmented. (C) Nuclear and Golgi complex surface and contours are calculated and overlaid. (D) The distance and angle between the nucleus border and every single pixel defining the Golgi complex. (E) The results are plotted using a box-plot graph where y is the distance of the nuclear border (0) to the Golgi complex in µm. The box illustrates the average spreading of the Golgi as well as the maximum and minimum distances between the nuclear envelope and the Golgi complex. All steps have been implemented as a Macro for ImageJ downloadable at http://rsb.info.nih.gov/ij/
Fig. S3. Nucleus position during linear migration. (A). Kymograph representation of the transmission channel of a single cell moving on a fibronectin line. The time is from left to right. (B) Kymograph representation of the nucleus channel of a single cell moving on a fibronectin line. (C). Kymograph representation of the Golgi channel of a single cell moving on a fibronectin line. (D) Schematic overlay of the three channels. (E) Overlay of the original channels.
Movie 1. Overview of Bsc1 cells migrating on 6-µm fibronectin lines. Frame rate: 1 frame per 15 minutes. Total length: 12 hours 15 minutes
Movie 2. The Golgi displacement is migration dependent. A Bsc1 cell transiently transfected with a nuclear marker (H2b-Hcred) and a Golgi marker (GalT-GFP) migrates on a 6-µm fibronectin line. Frame rate: 1 frame per 5 minutes. Total length: 8 hours
Movie 3. The Golgi complex is always maintained behind the nucleus. A Bsc1 cell transiently transfected with a nuclear marker (H2b-Hcred) and a Golgi marker (GalT-GFP) migrates on a 6-µm fibronectin line. Changes of direction: Frames 11-15 and 30-36. Frame rate: 1 frame per 15 minutes. Total length: 16 hours 45 minutes
Movie 4. The Golgi position is linked to the MTOC position. Sequence of a Bsc1 cell reorientation. A Bsc1 cell transiently transfected with a centrosomal marker (Centrin-Cherry) and a Golgi marker (GalT-GFP) migrates on a 6-µm fibronectin line. Left image: transmission image. Right image: GalT-GFP signal (Golgi matrix protein, green) and Centrin-Cherry (Centrosomal protein, red). Frame rate: 0.5 frames per hour.
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