Topotecan

The chemotherapeutic agents used included doxorubicin (Fisher Scientific, Waltham, MA, USA), paclitaxel (LKT Laboratories Inc.. St. Paul, MN, USA) and topotecan (Toronto Research Chemicals, Toronto, Canada). The statin drugs used in the research include atorvastatin (Toronto Research Chemicals Inc., Toronto, Canada), fluvastatin (Selleck Chemicals, Houston, TX, USA), lovastatin (EMD Millipore, Billerica, MA, USA), mevastatin (EMD Millipore, Billerica, MA, USA), pravastatin (EMD Millipore, Billerica, MA, USA) and simvastatin (EMD Millipore, Billerica, MA, USA). Mevalonolactone (Sigma-Aldrich, St. Louis, MO, USA) was dissolved in RPMI-1640 medium (Fisher Scientific, Waltham, MA, USA) and titrated with NaOH to a pH of 7.8 to form mevalonate. doxorubicin, fluvastatin and pravastatin were dissolved in distilled water. paclitaxel, topotecan, and atorvastatin were dissolved in methanol. lovastatin, mevastatin and simvastatin were dissolved in dimethyl sulfoxide (DMSO). The chemotherapeutic agents, statins, and mevalonate were aliquoted and stored at − 80 °C.

Reagents were obtained as follows: Navitoclax and A1210477 from MedChemExpress; venetoclax and S63845 from Chemietek; topotecan from Toronto Research Chemicals; olaparib, etoposide, and vincristine from Selleck Chemicals; 5-fluorouracil from Tokyo Chemical Industry; and CHAPS from Millipore-Sigma. Antibodies were purchased from the following suppliers: anti-PUMA (#sc-374223, 1:1000) and anti-actin (goat polyclonal, I-19, #sc-1615, 1:500) antibodies from Santa Cruz Biotechnology; anti-NOXA antibody from ENZO Life Sciences (#ALX-804-408-c100, 1:1000); anti-BCL2 antibody from DAKO (#M0887, 1:1000); anti-BAK antibody from Millipore (#06-536, 1:1000) and antibodies to BAX (#2772S, 1:1000), BCLX_L (#2764S, 1:1000), BIM (#2933S, 1:1000), MCL1 (#4572S, 1:1000), caspase-3 (#9662, 1:1000), caspase-9 (#9502, 1:1000), and tubulin (#2148, 1:1000) from Cell Signaling Technology. Rat anti-BID antibody was a kind gift from David Huang (Walter & Eliza Hall Institute, Melbourne, Australia).