Figure 2

CDDO-DFPA suppresses NF-κB signaling in LPS-activated BMDCs. (A) CDDO-DFPA impairs time-dependent induction of NF-κB in BMDCs expanded from the normal bone marrow of C57BL/6 mice that were pretreated in either the presence or absence of CDDO-DFPA (200 nM) for 1 hour before LPS (100 ng/ml) treatment for the indicated intervals. (B) BMDCs were pretreated in the presence or absence of CDDO-DFPA (100–400 nM) for 1 hour, followed by stimulation with LPS for 10 mins (100 ng/ml). The cytosolic, nuclear, and total cellular lysates were analyzed for phospho- and total NF-κB p65 and IκBα, β-actin, and Lamin B1 expression by Western blotting. All depicted blots are cropped and respective full-length blots are presented in the Supplementary Figure S6. (C) BMDCs were pretreated in the presence or absence of CDDO-DFPA (400 nM) for 1 hour, followed by stimulation with LPS for 10 mins (100 ng/ml). Cells were fixed and stained with NF-κB and DRAQ5 (nucleus), and images were acquired by confocal microscopy. All experiments were repeated a minimum of three times.