Fig. 3.

MX106/MX107 inhibits NF-κB activation by genotoxic agents. (A) MDA-MB-231 cells pretreated with dimethylsulfoxide or increasing doses of MX106 for 12 hours were exposed to Dox (2 µg/ml) for 2 hours as indicated. NF-κB activity was examined by EMSA. Whole cell extracts were analyzed by Western blotting with the indicated antibodies. (B) HEK293 cells were pretreated with dimethylsulfoxide, MX106 (3 µM), or MX107 (3 µM) for 12 hours. Cells were then exposed to IR (10 Gy) and harvested at 2 hours after treatment. Whole cell extracts were analyzed as in (A). (C) U2OS cells were treated with Dox (2 µg/ml) with or without MX106/MX107 pretreatment as in (B) and analyzed by immunoblotting as indicated. (D) HEK293 cells were treated with TNFα (10 ng/ml, 15 minutes) or VP16 (10 µM, 2 hours) in a setting similar to (B). Whole cell lysates were analyzed accordingly. (E) The transactivity of NF-κB was monitored with NF-κB Luciferase reporter in MDA-MB-231 cells after the indicated treatment. (F) The mRNA levels of IL-6, IL-8, and IκBα (NFKBIA) were analyzed by quantitative polymerase chain reaction in MDA-MB-231 cells treated with Dox (2 µg/ml), MX106 (3 µM), or MX107 (3 µM) alone or in combination as indicated. EMSA, Electrophoretic Mobility Shift Assay; RLU, relative light unit.