Rabbit anti pjnk thr 183 tyr 185

0.4-μM Transwells were bought from Costar Corp. (USA). Serum levels of alanine transaminase (ALT), aspartate transaminase (AST), lactic dehydrogenase (LDH) in patients were determined by routine laboratory tests in Anhui Provincial Hospital (Hefei, China). Kits for serum Hydroxyproline (Hyp) was bought from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Recombinant Human Wnt 5a was bought from R&D Systems (645-WN-010/CF). Viability of Hepatic stellate cell line (LX-2) cells was detected by Cyto Tox 96 Nonradioactive Cytotoxicity assay from Promega (USA). The primary antibodies were as following: rabbit anti-Jnk (#9252, Cell signaling), rabbit anti-pJnk (Thr 183/Tyr 185, #4668, Cell signaling), mouse anti-α-SMA (ab5694, Abcam), mouse anti-NFATc1 (sc-7294, Santa Cruz), rabbit anti-Collagen 1A1 (MA1-141, ThermoFisher Scientific, USA), mouse anti-β-actin (ab8227, Abcam). Horseradish peroxidase conjugated secondary antibodies were obtained from GE Healthcare (UK).

The RAW 264.7 macrophage cell line was obtained from ATCC (Manassas, VA, USA). Cells were incubated in Dulbecco’s modified Eagle’s Medium (DMEM; Invitrogen, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS; Invitrogen) and antibiotics (100 U/mL penicillin and 100 µg/mL streptomycin; GIBCO BRL, Grand Island, NY, USA) at 37 °C in a humidified incubator under 5% CO₂. L7Gn was purchased from NPBANK (National Institute for Korean Medicine Development, Gyeongsan, Korea). LPS and DMSO were purchased from Sigma-Aldrich (St. Louis, MO, USA). EZ-Cytox solution was from DAEIL lab (Seoul, Korea). TOPscript cDNA synthesis kit and 2X TOPsimple™ DyeMIX-nTaq were purchased from Enzynomics (Daejeon, Korea). Rabbit anti-inhibitor of κBα (IκBα), rabbit anti-phospho (p)-IκBα (Ser-32/36), mouse anti-JNK, mouse anti-p38, rabbit anti-iNOS, goat anti-COX-2, rabbit anti-IL-1 receptor-associated kinase 1 (IRAK1), mouse anti-α-tubulin, and rabbit anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Rabbit anti-p-p38 (Thr-180/Tyr-182), mouse anti-p-ERK (Thr-202/Tyr-204), rabbit anti-ERK, rabbit anti-p-JNK (Thr-183/Tyr-185), rabbit anti-transforming growth factor-β-activated kinase 1 (TAK1), and rabbit anti-p-TAK1 (Thr-184/Tyr-187) antibodies were purchased from Cell Signaling Technology Inc. (Danvers, MA, USA).