FIGURE 6.

Protein acetylation is not involved in IκBζ-mediated Lcn2 activation. A and B, effect of anacardic acid (A) and nicotinamide (B) on IκBζ-mediated Lcn2 activation. p50-/IκBζ-deficient MEFs were transfected with the following plasmids: the luciferase reporter plasmid pGL3-Basic containing the upstream region of Lcn2 (−1031/+54), the internal control plasmid pRL-TK, and the pcDNA3 vector or pcDNA3 for expression of FLAG-IκBζ and HA-p50. Cells were incubated for 12 h with the indicated concentrations of the histone acetyl transferase inhibitor anacardic acid (A) or the HDAC inhibitor nicotinamide (B), and luciferase activities were determined as described under “Experimental Procedures.” Results are means ± S.D. for three independent transfections. C, effect of the HDAC inhibitor TSA on IκBζ. HEK293T cells expressing FLAG-IκBζ were incubated for 6 h with 2 μm TSA. The cell lysates were applied to immunoprecipitation (IP) with anti-FLAG antibody. Precipitated FLAG-IκBζ as well as acid extracted histones in the nuclear extracts were analyzed by immunoblot with the indicated antibodies or by staining with CBB. Positions for marker proteins are indicated in kilodaltons.