Extended Data Fig. 4: Kdm5d deletion suppresses tumour cell metastasis.
From: Histone demethylase KDM5D upregulation drives sex differences in colon cancer
a, qPCR of Kdm5d in L2-3 and two independent Kdm5d knockout cell lines (‘KO1’ and ‘KO2’) derived from L2-3. Relative expression was calculated by normalizing to ‘L2-3’ (n = 3 wells per group). b, Proliferation assay of 3346, L2-3 and KO1 (n = 3 wells per group; error bars indicate s.d.; two-tailed Mann-Whitney test; no adjustment for multiple comparisons). c, Representative images of migration and invasion assay. d, Quantitation of c (n = 3 wells per group). e, Growth curves of subcutaneous tumours in male nude mice (two-tailed Mann-Whitney test; no adjustment for multiple comparisons). f, Liver metastases detected by luminescence in male nude mice with intrasplenically injected male iKAP cells with a luciferase reporter (n = 5 mice per group). g, Quantitation of luminescence signals in f (n = 5 mice per group). h, Kaplan-Meier OS analysis of mice in f (n = 5 mice per group; log-rank (Mantel-Cox) test). i, Representative liver tissues with metastatic tumours from an independent experimental repeat of f–h (n = 3 livers per group). j, Number of gross surface metastasis in livers in i. Cells injected into spleen metastasize to liver through portal vein. Surrounding the blood vessel, the cells form initial liver metastasis, followed by whole liver spread. The formation of liver surface metastasis indicates higher metastatic potential of the cells and more aggressive disease. k, Percentage of histologically metastatic lesion area in all liver lobes in each mouse. l, Representative histology of liver metastasis (representing three tissues per group). m, qPCR of KDM5D in 2806, a cell line derived from female iKAP distal primary CRC, and 2806 with overexpression of WT or enzymatic-dead (‘ED’) human KDM5D. Relative expression was calculated by normalizing to ‘hKDM5DWT OE’ (n = 3 wells per group). n, Representative images of migration and invasion assays of cell lines in m. o, Quantitation of n (n = 3 wells per group). p, Liver metastases detected by luminescence in female nude mice with intrasplenically injected female iKAP cells with a luciferase reporter (n = 3 mice per group). q, Quantitation of luminescence signals in p (n = 3 mice per group, except for n = 2 mice for hKDM5DWT OE group at 27 days). r, Liver metastatic tumours from mice in p. For all bar/dot plots (a,d,g,j,k,m,o,q), data are shown as mean value ± s.d.; P was derived with two-tailed unpaired t test; no adjustment for multiple comparisons.
