Fig. 5

The OTUD6A-CDC6 axis promotes tumour growth. a, The protein expression levels of OTUD6A in BCa cells and uroepithelial SV-HUC-1 cells were determined by Western blotting. b, c, CDC6 and OTUD6A expression levels in the indicated T24 (b) and UMUC3 (c) cells were measured by Western blotting. d, e, The proliferation of OTUD6A knockdown and control T24 cells was examined by CCK8 assays (d) and EdU incorporation assays (e). f, g, The effect of OTUD6A overexpression on UMUC3 cell proliferation was examined by CCK8 assays (f) and EdU incorporation assays (g). h, Growth curves of the indicated T24 tumours are shown. Tumours were measured every 4 days. i, An image of subcutaneous tumours formed by the indicated T24 cells is shown. j, The indicated T24 tumours were weighed. k, Representative IHC images indicating Ki-67, CDC6 and OTUD6A expression in the indicated T24 tumours are shown. Scale bars, 50 μm (left) and 20 μm (right). l, m, CCK8 assays (l) and EdU incorporation assays (m) were used to examine the proliferation of the indicated T24 cells. n, Chromatin-bound proteins (CBP) were extracted from the indicated T24 cells and analysed by Western blotting. o, Growth curves of the indicated T24 tumours are shown. Tumours were measured every 4 days. p, The indicated T24 tumours were weighed. q, EdU incorporation assays were used to examine the proliferation of the indicated T24 cells. r, Different histologic types in the indicated mouse bladders after 20 weeks of BBN treatment are shown (WT mice, n = 7; CKO mice, n = 6). s, The mouse bladders were weighed. t, Representative H&E staining images and IHC images indicating Ki-67, CDC6 and OTUD6A expression in the indicated mouse bladders. Scale bars, 100 μm (left) and 50 μm (right). All quantitative analyses were based on three independent experiments. The error bars indicate the SDs. *P < 0.05, **P < 0.01, ***P < 0.001, n.s. not significant, based on two-tailed Student’s t test