O bottom) are U-251MG, A-549, MCF-7, Hep-G2, A-431 and HeLa in the left column, and CaCo2, PC-3, RT-4, Hek-293, and U-20S in the right. doi:10.1371/journal.pone.0050292.gintact cells across different cell lines. Methods such as electron microscopy can image intact cells, but have interference from other cell components [11]. More invasive methods of preparation such as extraction of the microtubule network can allow electron microscopy to generate traceable images, but are no longer representative of intact cells [12]. Fluorescence microscopy, on the other hand, can be used to obtain information about proteins atmonomer-level resolution of localization without interference from other cell components in intact cells with high-throughput data. One reason for studying microtubule distributions across cell lines is to begin to search for explanations of how expression of microtubule-associated proteins (MAPs) may account for any differences observed. The expression levels of many proteins vary across cell lines [13], and there are cell-specific proteins thatComparison of Microtubule DistributionsFigure 6. Comparison of the bivariate distributions of the estimated model parameters of the eleven cell lines. The ellipses are centered at the bivariate means of the two parameters and contain about 67 to 80 of the cells for a particular cell line (at most 1.5 standard deviations from the means). doi:10.1371/journal.pone.0050292.gFigure 7. Hierarchical clustering trees of eleven cell lines. The trees were built on the pairwise Hotelling’s T2 statistics from (A) the testing of the bivariate distributions of the estimated number of microtubules and mean length and (B) from the testing of the bivariate distributions of the first two principal components of the multivariate features ASA-404 biological activity computed from the real images. doi:10.1371/journal.pone.0050292.gComparison of Microtubule DistributionsTable 3. Statistical tests of the model parameters and the features Dorsomorphin (dihydrochloride) site between cell lines.p-valuesU-251MG (94) CaCo2(77) A-549(66) PC-3(110) MCF-7(54) RT-4(38) Hep-G2(51) Hek-293(70) A-431(112) U-2OS(114) HeLa(35)U-251MG NA 1 0.077 1 1 0.11 5.7e-4* 4.3e-3* 1.5e-4* 2.6e-7* 0*CaCo2 0* NA 1 1 1 0.030* 1 0.92 8.7e-6* 1.1e-5* 0*A-549 0* 1 NA 1 1 5.4e-4* 1 1 2.7e-9* 1.9e-4* 0*PC-3 0* 1 1 NA 1 0.067 2.0e-3* 0.26 0.012* 0.12 0*MCF-7 0* 0.86 0.012* 0.62 NA 1 0.081 0.12 0.059 4.1e-3* 0*RT-4 0* 0.045* 0.32 1 4.9e-5* NA 1.0e-4* 2.0e-9* 7.1e-3* 8.6e-6* 0*Hep-G2 6.1e-13* 6.3e-6* 0.12 7.6e-4* 9.2e-12* 7.3e-5* NA 1 0* 0* 0*Hek-293 1.1e-10* 5.5e-3* 1 1 3.1e-6* 1 0.020* NA 0* 2.9e-11* 0*A-431 5.8e-6* 0* 0* 0* 0* 0* 0* 0* NA 1 0*U-2OS 9.8e-10* 0* 7.3e-12* 0* 0* 0.029* 0* 4.1e-7* 7.0e-6* NA 0*HeLa 0* 0* 0* 0* 0* 3.1e-5* 0* 0* 0* 6.1e-13* NAThe lower triangular part of the table is for the testing of equality of the bivariate 1527786 mean of the distribution of two estimated microtubule parameters (number of microtubules and mean of length) between cell lines using Hotelling’s T2 test. The upper part (Italic) is for testing of the equality of the bivariate mean of the distribution of the first two principal components (learned from and representing the multivariate distribution of features on real cells). The rows and columns of the table are sorted according to the tree from Figure 7 (A). The p-values are adjusted according to the family-wise Bonferroni correction for multiple testing. The “*” denotes cell lines which differ at significance level alpha = 0.05. The number in the parenthesis of the.O bottom) are U-251MG, A-549, MCF-7, Hep-G2, A-431 and HeLa in the left column, and CaCo2, PC-3, RT-4, Hek-293, and U-20S in the right. doi:10.1371/journal.pone.0050292.gintact cells across different cell lines. Methods such as electron microscopy can image intact cells, but have interference from other cell components [11]. More invasive methods of preparation such as extraction of the microtubule network can allow electron microscopy to generate traceable images, but are no longer representative of intact cells [12]. Fluorescence microscopy, on the other hand, can be used to obtain information about proteins atmonomer-level resolution of localization without interference from other cell components in intact cells with high-throughput data. One reason for studying microtubule distributions across cell lines is to begin to search for explanations of how expression of microtubule-associated proteins (MAPs) may account for any differences observed. The expression levels of many proteins vary across cell lines [13], and there are cell-specific proteins thatComparison of Microtubule DistributionsFigure 6. Comparison of the bivariate distributions of the estimated model parameters of the eleven cell lines. The ellipses are centered at the bivariate means of the two parameters and contain about 67 to 80 of the cells for a particular cell line (at most 1.5 standard deviations from the means). doi:10.1371/journal.pone.0050292.gFigure 7. Hierarchical clustering trees of eleven cell lines. The trees were built on the pairwise Hotelling’s T2 statistics from (A) the testing of the bivariate distributions of the estimated number of microtubules and mean length and (B) from the testing of the bivariate distributions of the first two principal components of the multivariate features computed from the real images. doi:10.1371/journal.pone.0050292.gComparison of Microtubule DistributionsTable 3. Statistical tests of the model parameters and the features between cell lines.p-valuesU-251MG (94) CaCo2(77) A-549(66) PC-3(110) MCF-7(54) RT-4(38) Hep-G2(51) Hek-293(70) A-431(112) U-2OS(114) HeLa(35)U-251MG NA 1 0.077 1 1 0.11 5.7e-4* 4.3e-3* 1.5e-4* 2.6e-7* 0*CaCo2 0* NA 1 1 1 0.030* 1 0.92 8.7e-6* 1.1e-5* 0*A-549 0* 1 NA 1 1 5.4e-4* 1 1 2.7e-9* 1.9e-4* 0*PC-3 0* 1 1 NA 1 0.067 2.0e-3* 0.26 0.012* 0.12 0*MCF-7 0* 0.86 0.012* 0.62 NA 1 0.081 0.12 0.059 4.1e-3* 0*RT-4 0* 0.045* 0.32 1 4.9e-5* NA 1.0e-4* 2.0e-9* 7.1e-3* 8.6e-6* 0*Hep-G2 6.1e-13* 6.3e-6* 0.12 7.6e-4* 9.2e-12* 7.3e-5* NA 1 0* 0* 0*Hek-293 1.1e-10* 5.5e-3* 1 1 3.1e-6* 1 0.020* NA 0* 2.9e-11* 0*A-431 5.8e-6* 0* 0* 0* 0* 0* 0* 0* NA 1 0*U-2OS 9.8e-10* 0* 7.3e-12* 0* 0* 0.029* 0* 4.1e-7* 7.0e-6* NA 0*HeLa 0* 0* 0* 0* 0* 3.1e-5* 0* 0* 0* 6.1e-13* NAThe lower triangular part of the table is for the testing of equality of the bivariate 1527786 mean of the distribution of two estimated microtubule parameters (number of microtubules and mean of length) between cell lines using Hotelling’s T2 test. The upper part (Italic) is for testing of the equality of the bivariate mean of the distribution of the first two principal components (learned from and representing the multivariate distribution of features on real cells). The rows and columns of the table are sorted according to the tree from Figure 7 (A). The p-values are adjusted according to the family-wise Bonferroni correction for multiple testing. The “*” denotes cell lines which differ at significance level alpha = 0.05. The number in the parenthesis of the.
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