Phospho thr581 ser 376 ser360msk1

Serum-free keratinocyte growth medium (Medium 154S) containing low calcium (0.2 mM), bovine pituitary extract (BPE) and epidermal growth factor (EGF) were obtained from Kurabo (Tokyo, Japan). Human primary keratinocytes (HPKs) were purchased from Biopredic Intl (Saint-Gregoire, France). Human HaCaT keratinocytes [22 (link), 23 (link)] were kindly supplied by Dr. M. Furue (Kyushu University, School of Medicine, Department of Dermatology). Rabbit polyclonal anti-TGase 1 (NOV-1184-45: NB100-1844 Lot7E3) was purchased from Novus Biologicals (Littleton, CO, USA). Antibodies to phospho-ERK, ERK, phospho-p38, p38, phospho-JNK, JNK, phospho-AKT and AKT were purchased from Cell Signaling Technology (Danvers, MA, USA). Antibodies to phospho-Ser468/Ser276/Ser536NFκBp65, NFκBp65, phospho-Ser133CREB, CREB, c-Jun, phospho-Ser73c-Jun, c-Fos, ATF-2, phospho-Thr71ATF-2, MSK1 and phospho-Thr581/Ser 376/Ser360MSK1 were also purchased from Cell Signaling Technology. Antibodies to COX-2 and β-actin were obtained from Sigma-Aldrich Corp. (St. Louis, MO, USA). The NFκB activation inhibitor II (JSH-23), the MEK inhibitor (PD98059), the p38 inhibitor (SB203580), the JNK inhibitor (JNK inhibitor II) and H89 were purchased from Calbiochem (San Diego, CA, USA). Other chemicals including AX were obtained from Sigma-Aldrich Corp.

Antibodies to ERK, phospho-ERK, p38, phospho-p38, JNK, phospho-JNK, NFκBp65, phospho- Ser276/Ser536NFκBp65, CREB, phospho-Ser133CREB, c-Fos, ATF2, phospho-Thr71ATF2, MSK1 and phospho-Thr581/Ser 376/Ser360MSK1, JAK2 and phospho-Tyr221JAK2, STAT3 and phospho-Tyr705/Ser727STAT3 were purchased from Cell Signaling Technology (Danvers, MA, USA). Antibodies to β-actin and HYBID (Anti-KIAA1199) were obtained from Sigma-Aldrich Corp., St. Louis, MO, USA). The antibody to protein tyrosine phosphatase, nonreceptor type 9 (PTPMEG2/PTPN9) was purchased from Santa Cruz Biotechnology (Dallas, TX, USA). The NF-κB activation inhibitor II (JSH-23), the MEK inhibitor (U0126), and the STAT3 inhibitor were from Calbiochem (San Diego, CA, USA). The p38 inhibitor (SB239063) and the JNK inhibitor (JNK inhibitor II) were from Santa Cruz Biotechnology and Merck KGaA (Darmstadt, Germany), respectively. Okadaic acid and sodium orthovanadate were obtained from Calbiochem.