Fig. 6.

DUB3 causes chemotherapy resistance by upregulating NRF2 in HCT116 cells. a–d DUB3 causes paclitaxel resistance by upregulating NRF2. We established stable cell lines HCT116-vector, HCT116-NRF2 and HCT116-DUB3, knockout cell lines HCT116-NRF2^−/−-1 and HCT116-NRF2^−/−-2, and knockdown cell lines HCT116-shNC, HCT116-shDUB3-1, and HCT116-shDUB3-2. HCT116-NRF2^−/− was infected with lentivirus which contained NRF2 or DUB3 to get the stably rescue cell lines named as HCT116-NRF2^−/−-NRF2 and HCT116-NRF2^−/−-DUB3. These cell lines were seeded in a 96-well plate and treated with paclitaxel for 48 h at the indicated doses. Cell viability was measured by CCK-8 assays. e–h DUB3 promotes cell survival after paclitaxel treatment in HCT116 cells. Cells were seeded in a 6-well plate and treated with 5 nM and 10 nM paclitaxel for 48 h on the following day, after which the medium was replaced with fresh medium. Clones were stained with crystal violet and photographed 10 days later. i–j Rescue of DUB3 in NRF2-deficient cells cannot recover the resistance to camptothecin in HCT116 and RKO cells. The experiments were performed as for panel (a). Data were analyzed employing one-way ANOVA and presented as the mean ± SD (N = 3/group)