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. 2020 Mar 30;122(11):1673–1685. doi: 10.1038/s41416-020-0779-9

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© The Author(s), under exclusive licence to Cancer Research UK 2020

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Fig. 6 — a Frequency of CSCs in DLD-1 cells transduced with control non-silencing shRNA (shCtrl) or shCSN6 lentivirus, as measured by an LDA in vivo. Tumour morphology in each group. b Quantification of tumour initiation. c Tumour growth curve for mice subcutaneously injected with control or shCSN6 DLD-1 cells (1 × 10⁶ cells per mouse, six mice per group). d RNA was extracted from tumour tissues. Quantitative RT-PCR analysis was performed to measure the mRNA level of Aldh1a1. The data are presented as the means ± SDs. e Protein was extracted from the tumour tissues described in c and immunoblotted with the indicated antibodies. f Representative IHC staining for CSN6, TRIM21, OCT1 and ALDH1A1 in tumour tissues from mice. The scale bars represent 100 μm. g Representative IHC staining for CSN6, TRIM21 and ALDH1A1 in human CRC TMAs. Case 1 is representative of a patient with CSN6-high colon cancer. Case 2 is representative of a patient with non-CSN6-high colon cancer. The scale bars represent 100 μm. h Quantification of staining intensities from sections in g. CSN6 and TRIM21 show a negative correlation, while CSN6 and ALDH1A1 show a positive correlation. i Kaplan–Meier survival curves of overall survival time based on CSN6 and TRIM21 expression from TMA analysis. *P < 0.05 and ***P < 0.001.