Fig. 4. Activation of non-canonical NF-κB by DDX5 downregulation mediates NRF2 transcription.

A qRT-PCR of NRF2 mRNA using total RNA isolated from Huh7 cells transfected with siCtrl or siDDX5 (50 pM) and Huh7 DDX5^OE cells, treated ±SOR (10 µM) for 24 h. Data expressed as mean ± SEM from n = 3. *p < 0.05 by unpaired t-test. B Immunoblots of NRF2 using nuclear lysates from Huh7 and HepAD38 cell lines transfected with siCtrl or siDDX5 ±10 µM SOR for 24 h. For DDX5^OE, Dox-inducible-DDX5 cell lines grown with Dox (1.0 µg/ml) for 48 h and SOR for the last 24 h. A representative immunoblot is shown from n = 3. C RT-PCR quantification of NRF2 mRNA using total RNA isolated from Huh7 xenograft tumors, treated ±SOR. Data expressed as mean ± SEM from eight tumors described in Li et al. [7]. *p < 0.05 by unpaired t-test. D qRT-PCR of NRF2 mRNA using total RNA isolated from HepAD38 WT, DDX5^KD, and DDX5^OE cells, transfected with indicated siRNAs (50 pM each) and treated +/- SOR (10 µM) for 24 h. Data expressed as mean ± SEM from n = 3. **p < 0.01 by unpaired t-test. E ChIP assays of NRF2 and GAPDH promoters using IgG, NFKB2/p52 or RNA polymerase II (Pol II) antibodies, in WT and DDX5^KD HepAD38 cells. Data expressed as mean ± SEM from two independent experiments. **p < 0.01 by unpaired t-test. F Immunoblots of NRF2 using WT and DDX5^KO Huh7 cells treated ±SOR (10 μM) for 48 h. A representative immunoblot from n = 3.