Electrophoretic mobility shift assay with antibody supershift to identify the subunits of NF-κB in the kidney and liver of sickle mice exposed to H/R. Lanes 1 to 4, TNF-treated MVEC controls; lanes 5 to 9, sickle mouse kidneys after H/R; lanes 10 to 14, sickle mouse liver after H/R. The supershift assay shows the presence of the p65 (lanes 6 and 11) and p50 (lanes 8 and 13) subunits of NF-κB in the kidney and liver of the sickle mice after H/R. There is no retardation with antibodies to p52 and cRel, suggesting that these subunits are not involved in the H/R-induced activation of NF-κB. This experiment also confirms that the identified bands are indeed caused by the presence of NF-κB. MVEC indicates microvascular endothelial cells; TNF, tumor necrosis factor; H/R, hypoxia and reoxygenation; anti-p65/50/52/cRel, antibodies to these NF-κB subunits.