Supplementary Figure 8: Restoration of normal microtubule assembly rates and chromosomal stability accelerates CRC tumor growth in vitro and in vivo.

(a–c) Accelerated colony formation activity of SW620 and HCT116-CHK2^−/− cells after partial loss of CH-TOG or AURKA. Stable SW620 cell lines expressing control, CH-TOG shRNAs or AURKA shRNAs were subjected to colony formation assays in soft agar (2,000 cells per well) and colonies were counted. a: 2,000 cells per well, mean ± s.e.m., t-test, n = 3 independent experiments. (b) 5,000 cells per well, mean ± s.e.m., t-test, n = 4–6, 4–6 independent experiments. c: 2,000 cells per well, mean ± s.e.m., t-test, n = 4 independent experiments. (d–f) Growth of xenograft tumors derived from SW620 or HCT116 cells as indicated with or without partial repression of CH-TOG/CKAP5 or AURKA. The indicated cells were injected s.c. into both flanks of the mice and tumor volumes were determined over time. (d) The experiment shown is an extension to the experiment shown in Fig. 7c using an independent CH-TOG/CKAP5 shRNA expressing clone (mean ± s.e.m., n = 14–16 tumors as indicated). (e) The experiment shown is an independent extension to the experiment shown in Fig. 7d and additional clones were used (mean ± s.e.m., n = 9–10 tumors as indicated). (f) The experiment shown is an independent extension to the experiment shown in Fig. 7e and additional clones were used (mean ± s.e.m., n = 7–11 tumors as indicated). (g) Xenograft tumors maintain the partial repression of AURKA in vivo. Cells were re-isolated from three different SW620 xenograft tumors grown in three different mice and expressing control (clone 1) or AURKA shRNAs (clone 1). Cells were subjected to western blotting and protein levels of Aurora-A were detected. Representative western blots are shown. (h) Tumour cells re-isolated from xenograft tumors do not show gross alterations in cell cycle distribution. Cells re-isolated from the different xenograft tumors were subjected to FACS analyses and representative cell cycle profiles are shown. (i) The suppression of CIN in SW620 cells with reduced AURKA expression is maintained in xenograft tumors in vivo. Tumour cells re-isolated from xenograft tumors were subjected to karyotype analyses (50 cells per condition). Detailed data on karyotype analyses can be found in the Supplementary Table 1. Statistic source data for Supplementary Fig. 7 can be found in the Supplementary Table 2.