Phosho p38

Cells were obtained from the American Type Culture Collection (Manassas, VA, USA). All cells were cultured in appropriate medium containing 10% fetal bovine serum (FBS; Welgene, Gyeongsan, Korea), 1% penicillin-streptomycin, and 100 μg/mL gentamycin (Thermo Fisher Scientific, Waltham, MA, USA). All cell lines tested negative for mycoplasma contamination. The lines were authenticated by standard morphologic examination using microscopy. FTY720 was purchased from Echelon Biosciences (Salt Lake City, UT, USA). N-acetyl-l-cysteine was obtained from Calbiochem (San Diego, CA, USA). E64D and NBD-FTY720 were purchased from Cayman Chemical (Ann Arbor, MI, USA). TNF-α and z-VAD-fmk were purchased from R&D Systems (Minneapolis, MN, USA). PD98059, SB203580, SP600125, NecroX-5, and pepstatin A were obtained from Enzo Life Science (Farmington, NY, USA). Anti-PARP, anti-Cathepsin D, anti-LAMP1, and anti-Beclin 1 antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-phospho-ERK, phosho-p38, anti-phospho-JNK, and anti-Alix were purchased from Cell Signaling Technology (Beverly, MA, USA). The anti-ATG7 antibody was obtained from ProSci Inc. (Poway, CA, USA). The cycloheximide, H₂O₂, 3-methyladenine, actinomycin D, glutathione ethyl ester, and anti-actin antibody were obtained from Sigma (St. Louis, MO, USA).

Metformin, dexamethasone (DEX), rosiglitazone, 3-isobutyl-1-methylxanthie (IBMX), insulin, compound C, SP600125, and SB203580 were purchased from Sigma-Aldrich (St. Louis, MO, USA). MTT was obtained from HXBIO (Hangzhou, China). Oil Red O was purchased from Solarbio (Beijing, China). The triglyceride GPO-PAP enzymatic kit was obtained from Jiancheng Bioengineering institute (Nanjing, China). Phospho-AMPK, total AMPK, Phospho-ERK, total ERK, phosho-p38, total p38, phospho-JNK, total JNK, phospho-Akt, total Akt, FASN, PPARγ, C/EBPα, polyclonal β-Actin, and horseradish peroxidase-conjugated secondary antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA). RNA extraction kit was obtained from Aidlab Biotech (Beijing, China). The iScript circular DNA (cDNA) synthesis kit and synergy brands (SYBR) master mix were purchased from Bio-Rad (Hercules, CA, USA). The bicinchoninic acid (BCA) assay kit was obtained from Pierce (Rockford, IL, USA). The Western Lightening™ Plus-ECL (Enhanced Chemiluminescence) Substrate assay kit was obtained from Perkin-Elmer (Waltham, MA, USA).