Fig. 7.
Inhibition of NiCl2-induced endothelial MCP-1 expression by dominant-negative IKKβ.
HUVECs were transfected in a 1:3 ratio with pGreenLantern expressing GFPS65T and a plasmid expressing either empty expression vector or dominant-negative IKKβ (dn IKKβ). Cells were left unstimulated or exposed to 1.5 mM NiCl2 or 2 ng/mL TNFα for 12 hours. Successfully transfected cells, ie, cells labeled by green fluorescent protein, were analyzed for MCP-1 expression by flow cytometry as described in “Materials and methods.” Flow cytometry profiles of one representative experiment are shown. Open profiles represent isotype controls; shaded profiles indicate cells intracellularly labeled for MCP-1 expression.