3 4 5 dimethyl 2 thiazolyl 2 5 diphenyl 2htetrazolium bromide mtt reagent

Hirsuteine (purity: 98%) was obtained from Abphyto biotech (Chengdu, Sichuan, China). 3-(4,5-Dimethyl-2-thiazolyl)-2, 5-diphenyl-2Htetrazolium bromide (MTT) reagent was from Amresco (Solon, OH, USA). Propidium iodide (PI) was from Sigma-Aldrich (St. Louis, MO, USA). Annexin V-FITC/PI Apoptosis Detection Kit was from BD Biosciences. TRIzol reagent was purchased from Life Technologies (Carlsbad, CA, USA). Fetal bovine serum (FBS) was from Biological Industries (Kibbutz Beit-Haemek, Israel). The primary antibodies against cyclin B1 (1:1000, #4135), cdc2 (1:1000, #9116), p-Rb (1:1000, #8516), Caspase-3 (1:1000, #9662), β-actin (1:1000, #8457), anti-rabbit HRP-conjugated secondary antibodies (1:2000, #7074) and anti-mouse HRP-conjugated secondary antibodies (1:2000, #7076) were purchased from Cell Signaling Technology (Danvers, MA, USA). Antibodies against p53 (60283-1-lg), p21 (10355-1-AP), MDM2 (27883-1-AP), LC3 (14600-1-AP), p-p53 (Ser15) (67826-1-lg), p62 (18420-1-AP), Ki-67 (27309-1-AP), PUMA (55120-1-AP) were purchased from Proteintech (Wuhan, Hubei, China).

Digoxin was purchased from Aladdin (London, Ontario, Canada). Doxorubicin was obtained from Dalian Meilun Biological Product Factory (Dalian, Liaoning, China). Cisplatin and verapamil were purchased from Energy Chemical (Shanghai, China). Anti-CyclinD1, anti-Cdc2, anti-CyclinB1, anti-HIF1α, anti-p-Rb (phospho S780), and anti-β-actin antibodies, as well as anti-mouse and anti-rabbit horseradish peroxidase (HRP)-conjugated secondary antibodies, were obtained from Cell Signaling Technology (Danvers, MA, USA). Anti-p21 was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-p-Integrinβ1 (phospho T788+T789) was from Abcam (Cambridge, MA, USA). Anti-MMP2 and anti-MMP9 were obtained from Bioss (Beijing, China). Matrigel and Annexin V-fluorescein isothiocyanate/propidium iodide (Annexin V-FITC/PI) apoptosis-detection kits were from BD Biosciences (San Jose, CA, USA). Propidium iodide (PI) was from Sigma-Aldrich (St. Louis, MO, USA). Human VEGF-A kit was purchased from Jianglai biotech (Shanghai, China). 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) reagent was from Amresco (Solon, OH, USA).