Protein kinase C (PKC) isoforms are major regulators of cutaneous homeostasis and mediate inflammation in response to 12-O-tetradecanoylphorbol-13-acetate (TPA). We have previously reported that transgenic mice overexpressing PKCα in the skin exhibit severe intraepidermal neutrophilic inflammation and keratinocyte apoptosis when treated topically with TPA. Activation of PKCα increases the production of TNFα and the transcription of chemotactic factors (MIP-2, KC, S100A8/A9), vascular endothelial growth factor, and GM-CSF in K5-PKCα keratinocytes. In response to PKCα activation, NF-κB translocates to the nucleus and this is associated with IκB phosphorylation and degradation. Preventing IκB degradation reduces both the expression of inflammation-associated genes and chemoattractant release. To determine whether TNFα mediated NF-κB translocation and subsequent expression of proinflammatory factors, K5-PKCα mice were treated systemically with a dimeric soluble form of p75 TNFR (etanercept) or crossed with mice deficient for both TNFR isoforms, and keratinocytes were cultured in the presence of TNFα-neutralizing Abs. The in vivo treatment and TNFR deficiency did not prevent inflammation, and the in vitro treatment did not prevent NF-κB nuclear translocation after TPA. Together these results implicate PKCα as a regulator of a subset of cutaneous cytokines and chemokines responsible for intraepidermal inflammation independent of TNFα. PKCα inhibition may have therapeutic benefit in some human inflammatory skin disorders.