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Figure S2. TSA does not affect IκBa degradation. (A) RAW cells were treated with LPS and TSA for 30–60 min, and cell lysates were immunoblotted with antibody to IκBa. (B) TSA does not affect NF-κB nuclear translocation. RAW cells were treated with LPS and TSA for 0–60 min, and nuclear extracts were immunoblotted with antibody to p65. (C) TSA does not inhibit κB-binding activity. RAW cells were pretreated with TSA for 1 h, and then treated with LPS for 4 h. Nuclear extracts were incubated with radiolabeled oligonucleotides corresponding to binding elements in the NOS2 promoter, fractionated, and autoradiographed. Excess nonlabeled κB oligonucleotide was added to lane 5, and antibody to p50 was added to lane 6. (D) TSA does not decrease NOS2 promoter transactivation and does not decrease κB-dependent transactivation. RAW cells were stably transfected with a NOS2 promoter luciferase construct (top) or with a κB–luciferase reporter construct (bottom), treated with TSA or control, and stimulated with LPS, and the amount of luciferase was measured by a luminometer.