Keratinocyte Growth and Inflammation Modulation

Keratinocyte growth medium (KGM-SFM), pituitary gland extract and EGF were purchased from Gibco (Life Technologies, BRL, Grand Island, NY, USA). Arecoline, catalase, aspirin and 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) were obtained from Sigma (Sigma Chemical Company, St. Louis, MO, USA). ANE was prepared and weighed as previously described [25 (link), 32 (link)]. PGE₂ and 8-isoprostane (8-iso-PGF_2α) Enzyme-linked immunosorbant assay (ELISA) kits were obtained from Cayman Chemical Company (Ann Arbor, MI, USA). Human IL-1α ELISA kits were from PeproTech (Rocky Hill, NJ, USA), whereas ADAM17 and EGF ELISA kits were from R&D Systems (Minneapolis, MN, USA). GM6001, PD153035, AG490, pp2, U0126, α-naphthoflavone and ZnPP were obtained from Tocris or Cayman. Anti-EGF, anti-IL-1α and anti-TNFα neutralizing aby were obtained from PeproTech. Antibodies against COX-2, cdc2, ADAM9 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were from Santa Cruz, whereas antibodies for cytokeratin 5 and cytokeratin 14 were from GeneTex (GeneTex International Corporation, Global, Hsin-Chu, Taiwan). IRAK-1/4 inhibitor was from APExBIO (Houston, TX, USA).