Fig. 5

CRL4-COP1 E3 ligase complex ubiquitylates UTX for degradation. A IB analysis of WCL from LoVo and SW620 cells for indicated proteins. As indicated, cells were treated with DMSO, MG132, MG101, MLN4924, or Baf-A1 for 12 h before the cells were harvested. P27 was used as a positive control. B IB analysis of WCL from SW620 cells transfected with indicated siRNAs. C IB analysis of WCL from HEK293T cells co-transfected with Flag-UTX and siCOP1. D IB analysis of WCL derived from HEK293T cells co-transfected with various doses of Flag-COP1 and HA-UTX constructs and treated with MG132 as indicated. E IB analysis of WCL and IP from HEK293T cells transfected with the indicated HA-COP1 splice variants and Flag-UTX constructs. Cells were treated with MG132 (20 µM) for 12 h before they were harvested. F IB analysis of WCL and IP from HEK293T cells transfected with the indicated shRNAs, empty vector (EV), Flag-CUL4B, and HA-UTX constructs. Cells were treated with MG132 (20 µM) for 12 h before they were harvested. G IB analysis of WCL from different colon cancer cell lines. H IB analysis of WCL from HCT116 and LoVo cells transfected with siCOP1 or NC. I IB analysis of WCL from SW620 cells infected with lentiviruses expressing EV or Flag-COP1. J qPCR analysis of UTX expression in indicated CRC cell lines transfected with siCOP1 or NC or Flag-COP1 or EV. K IB analysis of WCL and IP from HEK293T cells transfected with the indicated plasmids in the presence of MG132 (20 µM) for 12 h before they were harvested. L Illustration of the isoforms of UTX WT and different VP mutants. M IB analysis of WCL from HEK293T cells co-transfected with Flag-COP1 and WT or mutant HA-UTX. Data information: In (J), data are presented as mean ± SEM (two-tailed Student’s t-test). ns, non-significance, ^***P < 0.001