5 5 dimethyl 1 pyrroline n oxide dmpo 2 7 dichlorodihydrofluorescein diacetate dcf da

Galangin was purchased from Santa Cruz Biotechnology Inc. (Dallas, TX, USA). [3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium] bromide (MTT), 1,1-diphenyl-2-picrylhydrazyl (DPPH), N-acetyl cysteine (NAC), 5,5-dimethyl-1-pyrroline-N oxide (DMPO), 2′,7′-dichlorodihydrofluorescein diacetate (DCF-DA), and Hoechst 33342 were purchased from Sigma-Aldrich Inc. (St. Louis, MO, USA). 5,5′,6,6′-Tetrachloro-1,1′, 3,3′-tetraethyl-benzimidazolylcarbocyanine iodide (JC-1) was purchased from Invitrogen (Carlsbad, CA, USA). Primary antibodies for Bax and Bcl-2 were purchased from Santa Cruz Biotechnology Inc. (Dallas, TX, USA). Primary antibodies for caspase-3 and caspase-9 were purchased from Cell Signaling Technology (Danvers, MA, USA). All other chemicals and reagents were of analytical grade.

Shikonin (5,8-dihydroxy-2-(1-hydroxy-4-methylpent-3-enyl)naphthalene-1,4-dione, Fig. 1A) was obtained from Cayman Chemical Company (Ann Arbor, MI, USA). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), Hoechst 33342, trypan blue solution, 5,5-dimethyl-1-pyrroline-N-oxide (DMPO), 2’,7’-dichlorodihydrofluorescein diacetate (DCF-DA), and anti-actin antibody were obtained from Sigma-Aldrich (St. Louis, MO, USA). 5,5’,6,6’-Tetrachloro-1,1’,3,3’-tetraethylbenzimidazolocarbocyanine iodide (JC-1) was obtained from Invitrogen (Thermo Fisher Scientific, Inc., Waltham, MA, USA). Anti-phospho-JNK, anti-JNK, anti-phospho-p38, and anti-caspase-3 antibodies along with SB203580 (p38 inhibitor), and U0126 (ERK inhibitor) were obtained from Cell Signaling Technology, Inc (Danvers, MA, USA). Anti-Bax, anti-Bcl-2, anti-caspase-9, anti-cytochrome c oxidase subunit 4 (COX4), anti-phospho-ERK, anti-ERK, anti-p38, anti-E-cadherin, and anti-N-cadherin antibodies were supplied by Santa Cruz Biotechnology, Inc (Dallas, TX, USA). The JNK inhibitor SP600125 was provided by Selleck Chemicals (Houston, TX, USA). All other chemicals and reagents used were of analytical grade.