TNF-α-regulated expression of adhesion molecules ICAM-1 and E-selectin dependence on the IKK2/NF-κB pathway. To control for successful activation of the endothelial cells later to be processed for microarray analysis as well as for functionality of the dominant-negative IKK2 mutant, flow cytometry was performed in parallel experiments. Briefly, wild-type primary endothelial cells (HUVECs, top) or cells infected with parental vector as control (middle) or with retrovirus carrying a dominant-negative mutant of IKK2 (bottom) were exposed to medium or 2 ng/mL TNF-α. Cells were immunostained for ICAM-1 (bold profiles), E-selectin (dotted profiles), or incubated with IgG isotype control (thin profiles). Successful infection of HUVECs with the retrovirus was confirmed by coexpression of the GFP gene.