Baohuoside 1

The human glioma cell line (U251) is provided by the Cell Bank of Chinese Academy of Sciences (Shanghai, China). U251 cells were maintained in Dulbecco’s Modified Eagle Medium (DMEM, Invitrogen, California, USA) supplemented with 5% fetal bovine serum (FBS, Invitrogen), 100 μg/mL streptomycin and 100 U/mL penicillin (Invitrogen). The doubling time of U251 cells is about 23 h. The cells were kept at 37 °C and 5% CO₂, passaged every 2–3 days. Baohuoside I was purchased from YuanYe Biotechnology (Shanghai, China) and was dissolved in DMSO as 100 mM. The FITC Annexin V Apoptosis Detection Kit was purchased from BD Pharmingen™ (556547). The Cell Counting Kit-8 (CCK-8) assay was purchased from biosharp (BS350B).

Baohuoside I was purchased from YuanYe biotechnology (Shanghai, China). Baohuoside I was dissolved in DMSO at a final concentration of 100 mM. Compound C (CC; Sigma-Aldrich, Missouri, US) was dissolved in DMSO as 10mM. The Annexin V-FITC Apoptosis kit was purchased from BestBio Company (Shanghai, China). The Cell Counting Kit-8 (CCK-8) assay was purchased from BestBio Company (Shanghai, China). The antibodies against mTOR (catalog no. ab2732), p62 (catalog no. ab155686) and caspase-3 (catalog no. ab2302) were purchased from Abcam. The antibodies against S6K1 (catalog no. CST 9202), phosphorylated (p)-S6K1 (catalog no. CST 9204S), AMPK (catalog no. CST 2532S), p-AMPKα1 (catalog no. CST 2537), p-mTOR (catalog no. CST 5536S), LC3A/B (catalog no. CST 12741), caspase 8 (catalog no. CST 4790) and Bax (catalog no. CST 5023S) were purchased from Cell Signaling Technology, Inc. The antibody against Bcl2 (catalog no. 12789-1-AP) and Cora Lite 488 conjugated Affinipure second antibody (catalog no. SA00013-2) was purchased from ProteinTech Group, Inc. The GAPDH antibody (catalog no. AP0063) was purchased from Bioworld Technology, Inc. The Ki67 antibody (catalog no. AF0198) was purchased from Affinity Biosciences, Inc.