Mouse anti human cox 2

Keratinocyte growth medium (KGM-SFM), pituitary gland extract and epidermal growth factor (EGF) etc. were from Gibco (Life Technologies, BRL, Grand Island, NY, USA). Catechol, arecoline, PD153035, manumycin A, pp2, curcumin, catalase, α-naphthoflavone, Zn-protoporphyrin dicoumarol 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). PGE₂ and PGF_2α ELISA kits were purchased from Cayman Chemical Company (Ann Arbor, MI, USA). Reagents for reverse transcription (RT) and polymerase chain reaction (PCR) were purchased from Invitrogen (Invitrogen Corporation, Carlsbad, CA, USA). Total RNA isolation kits were from Macherey-Nagel (Macherey-Nagel Inc., Easton, PA, USA). ANE and PBL extract were prepared and weighed as previously [12] (link), [15] (link). Hydroxychavicol (HC) was synthesized and characterized as before [16] . Specific PCR primer sets for COX-2, CYP450 isoforms, hemeoxygenase-1 (HO-1), cyclinB1, cdc25C, cdc2, keratin 5, keratin 14 and β-actin were synthesized by Genemed Biotechnologies, Inc. (San Francisco, CA, USA). Pathscan p-Src (Y416) and p-EGFR (Tyr845) ELISA kits were from Cell Signaling (Cell Signaling Technology Inc., MA, USA). EZ-Detect™ Ras activation assay kits were from Pierce (Rockford, IL, USA). Mouse anti-human COX-2 and anti-GAPDH antibody were from Santa Cruz.

C-Phycocyanin was extracted and purified in our lab, and dissolved in PBS as a stock solution and conserved at − 20 °C [23 (link)]. The cell cycle and apoptosis analysis kit and annexin V-FITC/PI apoptosis detection kit were purchased from Shanghai YEASEN Biotechnology Co., Ltd., Shanghai, China. The TUNEL detection kit was obtained from Beyotime Biotechnology, Shanghai, China. CCK8 and all other chemicals were of analytic grade and were also purchased from Beijing Solarbio Science & Technology, Beijing, China. Mouse anti-human COX-2, Cyclin D1, Cyclin E, CDK2, CDK4, p21, p27, Fas, cleaved-caspase 3, pro-caspase 3, ERK1/2, p-ERK1/2, JNK, p-JNK, p38 MAPK, p-p38 MAPK, AKT, and p-AKT monoclonal antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies against β-actin and all the second antibodies were purchased from Sigma-Aldrich.