Figure 2

Visualizing TNFα-induced NF-kappaB translocation. MEF cells were treated with 10 ng/ml of TNFα for 15 min and processed by the REAP method for western blotting, or were fixed for indirect immunofluorescence, staining for NF-kB (red) and DNA (blue). Panel A shows that staining for NF-kB was primarily cytoplasmic in the absence of stimulation (left panel) and that a significant amount of NF-kB staining appeared in the nucleus after TNFα treatment (right panel). Panel B shows whole cell (W), cytoplasmic (C) and nuclear (N) fractions prepared by REAP and blotted for NFkB, hnRNP (nuclear marker) and α-tublin (cytoplasmic marker).