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Figure 1—figure supplement 3. For unbiased assessment of co-localization, custom software was developed for automated detection of foci in confocal stacks and calculation of 3D co-localization in foci (see Materials and methods).(A) Left panel shows focus detection in the current slice (red circle) and in the adjacent slice (orange circle) of a confocal stack. Right panel shows voxel intensity of the identified foci after watershedding. (B) Plot of the co-localization index (see Materials and methods) of FP fusions to indicated proteins relative to GFP-Ktu after automated identification of foci. (C) Plot of co-localization index for the same data after computational randomization of foci position. Reduction in co-localization of all tested proteins after randomization was extremely significant (p<2.2×10−16), suggesting that the colocaliztion patterns in B did not occur by chance. Image published in: Huizar RL et al. (2018) © 2018, Huizar et al. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View

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