5 fluorouracil

Olaparib (Catalog# A4154), cisplatin (Catalog# A8321), carboplatin (Catalog# A2171), and 5-fluorouracil (Catalog# A4071) were all purchased from APExBIO company. UPF 1096 (Catalog# S8038), NMS-P118(Catalog# S8363), stenoparib (E7449) (Catalog# S8419), niraparib (Catalog# S2741), rucaparib (Catalog# S4948), and veliparib (ABT-888) (Catalog# S1004) were all purchased from Selleckchem.
cisplatin (Catalog# A10221) was purchased from AdooQ Bioscience. ART-558(Catalog# HY-141520), DNA-PK inhibitors PIK-75 hydrochloride (Catalog# HY-13281), Nedisertib (Catalog# HY-101570) and AZD-7648 (Catalog# HY-111783), RAD51 Inhibitor B02 (Catalog# HY-101462), and Olaparib (for in vivo experiment: Catalog# HY-10162) were all purchased from MCE (MedChem Express). ART-812 was synthetized by Dr. Wayne Childers at Temple University School of Pharmacy. All compounds were dissolved, aliquoted, and stored following the manufacturer’s instructions.

CD133⁺cells transfected with XRCC1 shRNA or negative control shRNA were cultured in 96 well plates (1 × 10⁴ cells/well) overnight. On the second day, the cells were treated with 5-Fluorouracil (5-FU, final concentration of 0.1 mg/L) (APExBIO, United States) (Paschall et al., 2016 (link)). After 72 h, CCK-8 solution was added to each well, and the cultures were incubated at 37°C for 4 h. Then, the cultures were detected by use of a microplate reader.
CD133⁺cells transfected with XRCC1 shRNA or negative control shRNA were cultured in 6 well plates (2 × 10⁵ cells/well) overnight. On the second day, the cells were treated with 5-Fluorouracil (5-FU, final concentration of 0.1 mg/L). After 72 h, cells were harvested to assess cell apoptosis by flow cytometry as above.

Cells were cultured in 96 well plates at a density of 1 × 10⁴ cells/well overnight. Then, these cells were under the treatment of 5-Fluorouracil (5-Fu, final concentration of 0.1 mg/L) (APExBIO, United States) for 72h 33 (link). Following, each well was added CCK-8 solution and then incubated at 37 ℃ for 4 h. Lastly, a microplate reader was applied to determine the absorbance.
Cells were cultured in 6 well plates at a density of 2 × 10⁵ cells/well overnight. Next, these cells underwent the disposal of 5-Fluorouracil (5-Fu, final concentration of 0.1 mg/L) for 72h. Then, cells were collected to evaluated cell apoptosis by flow cytometry as above description.