Rottlerin Inhibits ROS Formation and Prevents NF𝜅B Activation in MCF-7 and HT-29 Cells

Figure 2

Rottlerin inhibits TNF𝛼-induced NF-𝜅B/DNA binding activities in MCF-7 and HT-29 cells. Cells were pretreated with Rottlerin (20 μM) or with Vitamin E (50 μM) with subsequent stimulation with TNFα (10 ng/mL) for 4 hr. Nuclear extracts (20 μg) were used to determine NF-B DNA binding activities by electrophoretic mobility shift assay (EMSA) using DIG-labled oligonucleotide containing NF𝜅B element. The unlabeled NF𝜅B consensus oligomer was used as the specific competitor and unlabeled NF-L6 consensus oligomer as the nonspecific competitor. Bottom panel shows the bands quantification of a representative blot. Complexes were visualized by chemiluminescence. Lanes: C, control; R, Rottlerin; T, TNF𝛼; R+T; Rottlerin and TNF𝛼; V, Vitamin E; V+T, Vitamin E and TNF𝛼.