Fig. 5: Trib1 suppresses Nrf2 activity during liver regeneration.

A An ARE reporter was transfected into HepG2 cells with or without increasing doses of Trib1 followed by treatment with HGF (20 ng/ml). Luciferase activities were normalized by protein concentration and GFP fluorescence. N = 3 biological replicates. B A schematic diagram of the Nrf2 activity assay. C C57B6/L mice were injected with adenovirus carrying shRNA targeting Trib1 or an empty vector. GW3965 was administered by oral gavage 2 days prior to 2/3 partial hepatectomy. The mice were sacrificed 48 h after the surgery and liver tissues were assayed for Nrf2 activity. N = 5 mice for each group. D C57B6/L mice were injected with adenovirus carrying a Trib1 vector or an empty vector prior to 2/3 partial hepatectomy. The mice were sacrificed 48 h after the surgery and liver tissues were assayed for Nrf2 activity. N = 5 mice for each group. E Primary murine hepatocytes were transfected with siRNA targeting Trib1 or scrambled siRNA (SCR) followed by treatment with HGF (20 ng/ml) and/or GW3965 (2 μM) for 24 h. Nrf2 activity was assayed as described in Methods. N = 3 biological replicates. F Primary murine hepatocytes were transduced with indicated adenovirus followed by treatment with HGF (20 ng/ml) for 24 h. Nrf2 activity was assayed as described in “Methods”. N = 3 biological replicates. G C57B6/L mice were injected with adenovirus carrying shRNA targeting Trib1 or an empty vector. GW3965 was administered by oral gavage 2 days prior to 2/3 partial hepatectomy. The mice were sacrificed 48 h after the surgery and ChIP assay was performed with anti-Nrf2 or IgG using liver tissues. N = 3 mice for each group. H C57B6/L mice were injected with adenovirus carrying a Trib1 vector or an empty vector prior to 2/3 partial hepatectomy. The mice were sacrificed 48 h and ChIP assay was performed with anti-Nrf2 or IgG using liver tissues. N = 3 mice for each group. I Primary murine hepatocytes were transfected with indicated siRNAs followed by treatment with HGF (20 ng/ml) and GW3965 (2 μM) for 24 h. ChIP assay was performed with anti-Nrf2 or IgG. N = 3 biological replicates. J Primary murine hepatocytes were transduced with indicated adenovirus followed by treatment with HGF (20 ng/ml) for 24 h. ChIP assay was performed with anti-Nrf2 or IgG. N = 3 biological replicates. Data are expressed as mean ± S.D. *p < 0.05, one-way ANOVA with post-hoc Scheff´e.