Notch1 regulates nuclear retention of NFKB/P65 in response to TNFα.

<p>A. (<i>Right panel</i>) Hepatocytes were treated with (+) or without (−) TNFα and cytoplasmic and nuclear fractions were extracted. (<i>Left panel</i>) hepatocytes were transfected with either non-specific siRNA or siRNA specific for NOTCH1 and treated with TNFα and cytoplasmic and nuclear fractions were extracted. Whole cell lysates, cytoplasmic and nuclear extracts were subjected to immunoblotting with anti-P65. B. HEPG2 cells were treated with (+) or without (−) TNFα and immunostained with anti-P65. The arrows indicate nuclear localization of P65. C. Quantification of the P65 immunostaining signal in HEPG2 cells (5B). The numbers represent the average fluorescence intensity of P65 (n = 100). D. HEPG2 cells were transfected with either non-specific siRNA or siRNA specific for NOTCH1 and treated with TNFα. The arrows indicate nuclear localization of P65. E. Quantification of the P65 immunostaining signal in HEPG2 cells (5D). The numbers represent the average fluorescence intensity of P65 (n = 100). The ‘*’ represents p value<0.05 obtained by a two-tailed t test.