Fig. 1.

DUB3 is a potential DUB of NRF2 and correlated expressed with NRF2 in colon cancer. a DUB3 has the strongest effect on NRF2 protein level among the top 10 DUBs. HEK293T cells were cotransfected with NRF2-Myc and individual top 10 DUBs-Flag overexpresiong vectors. Forty-eight hours after transfection, the cell lysates were analyzed by western blot with the indicated antibodies. NRF2-Myc protein expression was normalized to GAPDH. b DUB3 has the strongest effect on NRF2 transcriptional activity among the top 10 DUBs. HEK293T cells were cotransfected with ARE reporter firefly luciferase (100 ng), pRL-TK (10 ng) and the indicated amounts of DUBs. Reporter assays were performed 24 h after transfection, and the results are presented as the NRF2/TK luciferase activity. c Oncomine data mining analysis of NRF2 mRNA levels in three different datasets comparing normal tissues vs. CRC tissues. d The protein expression of NRF2 and DUB3 in 24 representative pairs of primary CRC (T) and adjacent non-tumor tissues (N). e The protein expression of NRF2 and DUB3 in the human embryonic kidney cell line HEK293T, the normal human colon cell line FHC and seven colorectal cell lines (HCT116, SW48, SW480, RKO, DLD1, LOVO, and HT29)