Epithelial inflammatory chemokine elaboration requires NFκB signaling. Elevation of KC, MIP-2, and VEGF-A protein expression at baseline “C” and 3 hours after TPA treatment “T” in supernatants conditioned by primary transgenic keratinocytes (A, B, and C, respectively) demonstrate the cell autonomous activity of epithelial HIF-1α gain of function. (D) Increased NFκB transcriptional activity in K14-HIF-1αDPM keratinocytes. Nontransgenic (NTG) and transgenic (DPM) keratinocytes were transiently transfected with a NFκB reporter construct. After 48 hours, cells were treated with DMSO or TPA for 3 hours, harvested, and assayed for luciferase activity. Values are expressed as relative light units per microgram total protein (D). IκBα^Ser32/34A (IκBα super-repressor, IκBαSR) NFκB transcriptional blockade markedly diminishes keratinocyte chemokine expression (KC, MIP-2, and TNF-α, panels E-G, respectively) but does not affect the HIF-1α target gene (VEGF-A, PlGF, and Glut-1, panels H-J, respectively) expression. RT-PCR analysis using total RNA extracted from a control adenovirus (A-cytomegalovirus) or IκBαSR adenoviral-transduced primary keratinocytes at baseline and 3 hours after TPA treatment. Error bars represent mean plus or minus SEM. Results are representative of 3 independent experiments (*P < .05, t test).