Supplementary Figure 1: The expression level of EB3-GFP does not affect microtubule plus end assembly rates whereas partial repression of CH-TOG/CKAP5 and low dose Taxol treatment in various CRC cell lines suppresses chromosome number variability without affecting normal cell cycle progression.

(a) The average relative expression level of EB3-GFP is similar in all CRC cell lines investigated in microtubule assembly rate measurements. The relative expression levels of EB3-GFP were determined by quantifying the overall GFP fluorescence intensities. The graph shows mean values ± s.d. (n = 20 cells from 3 independent experiments). (b) HCT116 cells with representative high or low EB3-GFP expression levels (see panel a) were used to measure microtubule assembly rates. Scatter dot plots show average growth rates based on measurements of 20 microtubules per cell (mean ± s.e.m., t-test, n = 6 cells from 2 independent experiments). No difference was seen in cells with low versus cells with high-level expression of EB3-GFP. (c) Representative western blots showing protein levels for ch-TOG and α-tubulin in single cell clones derived from the indicated CRC cell lines stably expressing control or CH-TOG/CKAP5 shRNAs. Relative ch-TOG protein levels were quantified. (d) Karyotype analyses using CEP-FISH. Single cell clones derivative from SW620 cells expressing control or CH-TOG/CKAP5 shRNA were grown for 30 generations and subsequently subjected to CEP-FISH analysis. The proportion of cells that deviate from the modal chromosome number of chromosome 7 and chromosome 15 was calculated (100 cells analysed per condition). (e) FACS analyses of CRC cells after treatment with low dose Taxol show no cell cycle impairment. Single cell clones derived from the different CRC cells lines were generated in the absence (DMSO) or presence of low dose Taxol (0.05–0.5 nM). Cells were grown for 30 generations and were then subjected to FACS analyses and representative cell cycle profiles are given. (f) Karyotype analyses using CEP-FISH. SW837, SW620 or SW480 single cell clones were grown in the presence or absence of Taxol for 30 generations and subsequently subjected to CEP-FISH analysis. The proportion of cells that deviate from the modal chromosome number of chromosome 7 and chromosome 15 were calculated (100 cells analysed per condition). Detailed data on karyotype analyses can be found in the Supplementary Table 1. Statistic source data for Supplementary Fig. 1 can be found in the Supplementary Table 2.