Figure 3.

Cytokine-mediated activation and nuclear translocation of NF-κB is not significantly influenced by PR. A, T47D-A18 cells were treated with vehicle (v) or 10 nm R5020 (R) in the presence or absence of cytokines (10 ng/ml IL-1β or 50 ng/ml TNFα) for 6 h. mRNA levels of IκBα were quantitated using qPCR. Results were expressed as fold induction over vehicle-treated cells. Whole-cell extracts (WCE) and cytoplasmic extracts (CE) were resolved by SDS-PAGE and blotted with indicated antibodies. B, T47D-A18 cells were treated as in A and harvested 2 or 6 h after treatment. Nuclear extracts (NE) and WCE were resolved by SDS-PAGE and blotted with indicated antibodies. Equal amount of protein was confirmed by using GAPDH, cytokeratin 18, and lamin A as loading controls.