Analysis of NF-κB activation in mifepristone-treated mice after PH. (a) EMSA analysis of NF-κB DNA binding. Nuclear extracts were prepared from livers of mifepristone-treated single- or double-transgenic mice killed at the indicated times after PH. Five micrograms of nuclear protein was incubated with labeled NF-κB probe as described in Methods. Lanes represent individual animals. (b) IκBα Western blot of liver nuclear protein. Ten micrograms of liver nuclear extract protein used for EMSA shown in a was used for IκBα immunoblot. (c) Coincubation assay. Nuclear extract protein from 2-hour PH samples used in a were incubated alone or together at the indicated ratios with labeled NF-κB probe. (d) Histological analysis of p65 nuclear translocation in mifepristone-treated double-transgenic mice killed without PH or 2 hours after PH. Original magnification, ×400.