Trichloroacetic acid tca

Defatted tree peony seed flour was provided by the Huarui Science and Technology Development Co. Ltd. (Heze, Shandong, China). Soybean oil was purchased from a local supermarket in Jinan, Shandong, China. Alcalase 2.4 L (2.0 × 10⁵ U/g) was purchased from Novo Co., Ltd. (Beijing, China). The 1-anilino-8-naphthalene-sulphonate (ANS), DPPH, chloroform, tween-40, β-carotene, linoleic acid, glutathione (GSH), and 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA) were purchased from Sigma Co., Ltd. (St. Louis, MO, USA). Dulbecco’s modified Eagle medium (DMEM) and fetal bovine serum (FBS) were purchased from Gibco (Grand Island, NY, USA). Penicillin–streptomycin, 0.25% trypsin-EDTA, and trichloroacetic acid (TCA) were purchased from Beijing Solarbio Science Technology Co., Ltd. (Beijing, China). The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenylthiazolium bromide (MTT) and LDH assay kits were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). SOD, MDA, CAT, and Bicinchoninic acid (BCA) assay kits were purchased from Beyotime Institute of Biotechnology (Shanghai, China). All other chemicals and reagents used were of analytical grade.

Tetracenomycin X (Tcm X) was isolated from the marine-derived actinomycete Saccharothrix sp. 10-10 by Professor Maoluo Gan at our institute. The molecular weight of Tcm X was 486 Da, and it was dissolved in dimethyl sulfoxide (DMSO). The anti-p53, anti-PARP, anti-Bcl-2, anti-Bax, anti-p38 MAPK, anti-phospho-p38 MAPK, anti-cJUN, anti-phospho-cJUN, anti-cyclin D1 and anti-CDK4 antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). The anti-LC3B antibody was obtained from Sigma (St. Louis, MO, USA). The anti-Human CD261 Azide Free (DR4) antibody was obtained from Diaclone (Besancon, France), and the DR5 antibody was purchased from ProSci (San Diego, CA, USA). β-Actin (6G3), β-tubulin and GAPDH (glyceraldehyde-3-phosphate dehydrogenase) (1C4) mouse monoclonal antibodies were obtained from AmeriBiopharma (Wulmington, DE, USA). For the second antibodies, peroxidase-conjugated affiniPure goat anti-mouse IgG (H + L) and anti-rabbit IgG (H + L) were obtained from ZSGB-BIO (Beijing, China). MG132, cycloheximide (CHX), SB203580 and SP600125 were purchased from Selleck.cn (Houston, TX, USA). Sulforhodamine B (SRB) was obtained from Sigma (St. Louis, MO, USA). Trichloroacetic acid (TCA) was obtained from Solarbio (Beijing, China).