Cell apoptosis and cycle kits

Dulbecco’s Modified Eagle Medium (DMEM) was purchased from Gibco (Waltham, MA, USA). Trypsin was purchased from Biological Industries (Kibbutz Beit-Haemek, Israel). MTT was purchased from Solarbio (Beijing, China). Dimethyl sulfoxide (DMSO) was purchased from Zhiyuan Chemical Reagent Co., Ltd. (Tianjin, China). Cell apoptosis and cycle kits were purchased from BD Biosciences (San Jose, CA, USA). The trans-well chambers were purchased from Corning (Corning, NY, USA). The BCA protein assay kit was purchased from beyotime (Shanghai, China). Other reagents and solvents were of analytical grade or commercially available and used without further purification. The CO₂ cell incubator was purchased from Thermo Fisher Scientific (Waltham, MA, USA). The allegraX-15R high-speed centrifuge was purchased from Beckman (Brea, CA, USA). The microplate reader was purchased from BioTek (Winooski, VE, USA). ACEA NovoCyte flow cytometry analyzer was purchased from BD Biosciences (USA). The Leica inverted fluorescence microscope was purchased from Leica (Wetzlar, Germany).

Ultraviolet-visible (UV/Vis) spectra were acquired using a UV-Vis spectrophotometer, the UV-5300 (Hitachi, Tokyo, Japan). Infrared (IR) spectra were measured on a Bruke Vertex 80 (KBr disks) infrared spectrometer. Optical rotations were recorded in methanol (MeOH) solution on an AUTOPOL1 polarimeter at 28 °C (Rudolph, Wilmington, MA, USA). CD spectra were obtained on a J-810 circular dichroism spectropolarimeter (JASCO Corp., J-810, Tokyo, Japan). Electro-spray ionization mass spectrometry (ESIMS) and high-resolution electrospray ionization mass spectrometry (HRESIMS) analyses were measured on a Waters Xevo TQS and a Waters Xevo G2S Q-TOF LC/MS, respectively. NMR spectra were recorded on a Bruker 600 MHz instrument, and tetramethylsilane (TMS) was used as an internal standard. Column chromatography was performed on silica gel (200−300 mesh, Qingdao Marine Chemical Co., Ltd., Qingdao, China) and Sephadex LH-20 (Amersham Biosciences, Uppsala, Sweden), respectively. HPLC analysis was performed on an Agilent 1260 with a C18 column (Agilent Extend-C18, 4.6 × 250 mm, 5 μm, 1 mL/min). HPLC separation was performed on Shimadzu Soviet production LC-16P with an ODS column (Silgreen C18AB, 10 × 250 mm, 5 μm, 180 Å, 3 mL/min). Cell apoptosis and cycle kits were purchased from BD Biosciences (San Jose, CA, USA).