Veliparib

The following compounds were used: entinostat (Biomol GmbH, M4693-15A.25), idasanutlin (TargetMol; T6254), olaparib (BIOZOL; S1060), ribociclib (supplier), selinexor (BioCat; T6106-TM), vinblastinesulfate (Selleckchem; S4505), doxorubicin in 0.9% NaCl (UKHD pharmacy), niraparib (Selleckchem; S2741), pamiparib (Selleckchem; S8592), veliparib (Selleckchem; S1004). The translational drug library for initial drug selection comprised 74 compounds [19 (link)].

All used cell lines were held at 37 °C and 5% CO₂ unless stated otherwise and authenticated by fingerprinting at the DSMZ (Braunschweig, Germany), cultivated as follows and split every 2–3 days: DAUDI (ACC-78), HG-3 (ACC-765), JEKO-1 (ACC-553), JVM-2 (ACC-12), JVM-13 (ACC-13), MINO (ACC-687), PGA-1 (ACC-766), RAMOS (ACC-603), REC-1 (ACC-584), VAL (ACC-586), WA-C3CD5+ (ACC-769) cells were grown in RPMI 1640 medium with 10% fetal calf serum (FCS) and MEC-1 (ACC-497) in Iscove’s MDM with 10% FCS. 293 T (ACC-635) cells were grown in Dulbecco’s MEM with 10% FCS and at 10% CO₂. Inhibition of PRMT5 enzymatic activity was achieved with the indicated doses of EPZ015666 (Sigma, SML1421) for 96 h. Additional treatments were performed with the following chemicals: Veliparib (40 µM, 48 h; Selleckchem; S1004), SAHA (1 µM, 48 h; Sigma; SML0061), JQ1 (5 µM, 48 h; Sigma; SML0974), Chaetocin (50 nM, 24 h; Sigma; C9492).

A TRPC6 inhibitor or PARP1 inhibitor was administered during I/R injury. Rats in the I/R group were divided into five groups (n = 3 rats per group) according to whether OAG (Sigma, USA), SKF96365 (Sigma, USA), or veliparib (Selleck) was administered before the I/R procedure. Similarly, rats in the I/R + Nec-1 group were also divided into three groups (n = 3 rats per group). The rats were sacrificed at different time points (6, 12, or 24 h after reperfusion), and their kidneys were harvested. Next, sections of kidney tissue were fixed in 10% buffered formalin for histological examination. The remaining kidney tissue from each rat was frozen in liquid nitrogen and stored at −80°C for further analysis.

For the dose–response relationship, chondrosarcoma cells were pre-treated with 2.5 µM, 10 µM, and 25 µM of the PARPi Olaparib or Veliparib, the ATMi Ku-55933, or the ATRi VE-821 (all Selleckchem, Houston, TX, USA). Afterwards, the cells were irradiated with 0 Gy (non-IR control), with a fractionated IR of 8 Gy X-ray on each of the three subsequent days, or with a total dose of 24 Gy. Cell viability was determined with the CellTiter-Glo^® cell viability assay (Promega Corporation, Madison, MI, USA) and normalized to the non-IR controls. Background reference values were derived from the culture media. Absorbance was measured with a LUMIstar™ microplate luminometer (BMG Labtech GmbH, Ortenberg, Germany) (mean ± SD; n = 3, performed in biological quadruplicates).
The xCELLigence RTCA-DP device (OLS, Bremen, Germany) was used to monitor cell proliferation in real-time. Cells were seeded after IR in electronic microtiter plates (E-Plate™, OLS) and measured for 120 h according to the manufacturer’s instructions. Cell density was measured in quadruplicate with a programmed signal detection every 20 min. Data acquisition and analyses were performed with RTCA software (version 1.2, OLS).