Electrophoretic mobility shift assay (EMSA) demonstrates that NF-κB is up-regulated in the lungs of transgenic NY-S mice and LPS-treated normal mice (18 hours after LPS injection) compared with normal lung controls.
Lung cell extracts were isolated from lung homogenates prepared from 3 NY-S mice, 3 normal control mice, and 2 LPS-treated controls. Lung cell extracts from 260 ng lung DNA were incubated with end-labeled32P-dsDNA containing a consensus murine NF-κB DNA binding sequence (underlined): 5′-AGTTGAGGGGACTTTCCCAGGC-3′. DNA-protein binding reactions were carried out with each extract and separated by electrophoresis. Radioactive bands corresponding to NF-κB–DNA complexes were quantified by phosphorescence densitometry. The figure shows the bands from the lungs of one LPS-treated normal mouse, 3 normal control mice, and 3 transgenic NY-S mice. The summary bar graph plots the mean ± SD lung NF-κB expression for each mouse group as a percentage of normal control mice (n = 3 for NY-S and normal control, n = 2 for LPS-treated control). *P ≤ .05; **P ≤ .01.
