Genipin

A reference standard of shanzhiside (purity 98%) was purchased from Shanghai Tauto Bio-tech Co., Ltd. (Shanghai, China). A reference standard of geniposide (purity 98%) was purchased from Shanghai ANPEL Laboratory Technologies Inc. (Shanghai, China). Reference standards of genipin-1-gentiobioside and genipin (purity 98%) were purchased from Dalian Meilun Biotechnology Co., Ltd. (Dalian, Liaoning, China). Paeoniflorin (purity 98%) was purchased from Shanghai Yuanye Bio-technology Co., Ltd. (Shanghai, China) and used as an internal standard (IS). GF extract was prepared in our lab and the chemical analysis was performed as our previous research (The proportions of shanzhiside, genipin-1-gentiobioside and geniposide were 0.06%, 3.75% and 12.45%, respectively) [27 ]. The herbal materials of SSP were purchased from Chengdu and authenticated by Dr. Luping Qin from the Second Military Medical University. Acetonitrile, methanol (analytical grade) and acetic acid (HPLC grade) were purchased from Merck Company (Darmstadt, Germany) and Tedia (Fairfield, USA), respectively. Deionized water (18.2 MΩ/cm) was generated in-house using a Milli-Q System from Millipore (Bedford, MA, USA).

Genipin, geniposide and geniposidic acid were purchased from Dalian Meilun Biology Technology Co., Ltd. 5-Fluorouracil (5-FU) was purchased from MilliporeSigma. The chemical reagents were dissolved in DMSO to prepare stock solutions (100 µM). Antibodies against cyclin-dependent kinase 2 (CDK2; cat. no. ab32147), cyclin A2 (cat. no. ab181591), cleaved caspase-3 (cat. no. ab32042), cleaved poly-ADP ribose polymerase (PARP; cat. no. ab32561), E-cadherin (cat. no. ab40772), MMP-2 (cat. no. ab92536), Beclin-1 (cat. no. ab210498), light chain 3 (LC3; cat. no. ab192890), 5'-AMP-activated protein kinase (AMPK; cat. no. ab207442), phosphorylated (p)-JNK1/2/3 (cat. no. ab124956) and JNK1/2/3 (cat. no. ab179461) were purchased from Abcam. Anti-p-AMPK (cat. no. 2535) antibody was obtained from Cell Signaling Technology, Inc. Anti-rabbit IgG (cat. no. KGAA35) and β-actin (cat. no. KGAA006) were purchased from Jiangsu KeyGen Biotech Co., Ltd. SP600125 (cat. no. T3109) and BML-275 (cat. no. T1977) was purchased from Shanghai Topscience Co., Ltd.