Tetrandrine

The erythroleukemia cell line HEL (human origin) was originally obtained from ATCC (HEL 92.1.7), and maintained mycoplasma free used in all experiments.19 (link),20 (link) The generation of inducible K562-fli1 cells is described elsewhere.20 (link) These cells were maintained in Dulbecco’s Modified Eagle Medium supplemented with HyClone 5% fetal bovine serum (GE Healthcare).
The HEL cells were treated with indicated drugs, used for cell counting/MTT assay (to determine the proliferation index) and for protein/mRNA extraction (for Western blotting and RT-qPCR). Famotidine was purchased from (APExBIO), Tetrandrine from (Solarbio, CN), EGC from (Selleckchem, CN) and Diacerein from (APExBIO).

Tetrandrine (purity ≥ 98%) was purchased from Aladdin Biochemical Technology Co. (Shanghai, China). Tetrandrine was dissolved in DMSO (Solarbio Science & Technology Co., Beijing, China) and was then added to the cell culture supernatant in appropriate proportions. FBS and tyrosine were obtained from Gibco (Carlsbad, CA, USA). Cell Counting Kit-8 (CCK-8), Hoechst 33258 and Annexin V-PI apoptosis reagents were purchased from Beyotime (Shanghai, China). The TRIzol reagent and qRT-PCR Kit were purchased from Invitrogen (Carlsbad, CA, USA). A cDNA Synthesis Kit was from Takara (Shiga, Japan). The antibodies against Bax, Bcl-2, Akt, p-Akt, and β-actin were purchased from Cell Signalling Technology, Inc. (Danvers, MA, USA). The HRP and FITC goat anti-rabbit antibodies were purchased from Wanleibio Co. (Shenyang, China). The relevant primers were synthesized by Rui Biotech Co. (Qingdao, China).