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Image acquisition and quantitation was done by using Volocity software.
Intensity measurement was done by using Volocity software.
These regions were measured in 3D by using Volocity software (Improvision).
Seventeen Z stacking images with 0.2 μm intervals were acquired and quantitation was done by using Volocity software.
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All 3-dimensional images taken by confocal microscope were reconstructed using Volocity software.
The inherent fluorescent intensity of the CAE-positive cells were quantitated using Volocity software (Improvision) by dividing the sum of fluorescence intensity per field by the number of cells within that field (n>125 cells).
Background noise of NADH autofluorescence was removed by fine filter (Kernel 3 × 3) using Volocity software.
For cilia length, the length of at least 100 cilia (as judged by acetylated α-tubulin staining) was measured using Volocity software.
For the distance between the basal bodies/centrioles, the distance between mother and daughter basal bodies/centrioles (as judged by brightness of centrin-1/GFP) was measured using Volocity software.
The number of Tia1 and Dcp1a objects in each microscopy field was calculated based on florescence intensity signal using Volocity software and was divided by the number of cells in each field.
All immunofluorescence was visualized by confocal microscopy (LSM 700), and images were processed using Volocity software (PerkinElmer Life Sciences).
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