Figure 1

CRBN overexpression inhibits TLR4-induced NF-κB activation. (a) 293/TLR4 cells were transfected with a mock or HA-CRBN vector together with a pBIIx-luc and Renilla luciferase vector. Twenty-four hours after transfection, cells were untreated or treated with LPS (200 ng/ml) for 6 h and then analyzed for luciferase activity. Results are expressed as the fold induction in luciferase activity relative to that in untreated cells. All error bars represent standard deviation (S.D.) of the mean from triplicate samples. ns, not significant; **P<0.01. (b) 293/TLR4 cells were transfected with a mock or HA-CRBN vector, treated with or without LPS (200 ng/ml) for 6 h, and then analyzed for p65-DNA binding activity using the manufacturer’s protocol. All error bars represent S.D. of the mean from triplicate samples. ns, not significant; *P<0.05. (c) THP-1 cells were transfected with mock or different concentrations of HA-CRBN vector together with the pBIIx-luc and Renilla luciferase vectors. Twenty-four hours after transfection, cells were untreated or treated with LPS (200 ng/ml) for 6 h and then analyzed for luciferase activity. Results are expressed as the fold induction in luciferase activity relative to that in untreated cells. All error bars represent S.D. of the mean from triplicate samples. ns, not significant; *P<0.05. (d,e) THP-1 cells were transfected with mock or different concentrations of HA-CRBN vector, treated with or without LPS (200 ng/ml) for 9 h, and production of IL-6 (d) and IL-1β (e) were analyzed by enzyme-linked immunosorbent assay (ELISA). All error bars represent S.D. of the mean from triplicate samples. *P<0.05