Kw2449

The murine macrophage-like cell line RAW 264.7 was obtained from the ATCC. The cells were cultured in Dulbecco’s modified Eagle’s medium (Gibco) supplemented with 10% (v/v) fetal bovine plasma (Gibco) with 100 U/ml penicillin, 100 μg/ml streptomycin (Gibco) and 100 μg/ml Primocin (In vivoGen) at 37°C in a humid atmosphere with 5% CO₂. Then, RAW 264.7 cells were reseeded in 96-well or 6-well culture plates at densities of 1×10⁵ or 1×10⁶ cells/well and used for follow-up experiments. RAW 264.7 cells were stimulated with LPS (1 μg/ml) to establish a cell inflammation model and LPS (1 μg/ml) plus zVAD (20 μM; APExBIO) to establish a necroptosis model, with or without KW2449 (1 μM) or RIPA56 (1 μM; MedChemExpress LLC). The cell supernatant was collected 24 h after drug addition, and the cells were collected at the corresponding time points for detection.

We studied 8 PKI with PLK4 cross-over potential: CFI-400945 (CAS#1338800-06-8, Cat#16850) (Cayman Chemical, Ann Arbor, MI, USA) [55 (link)]; CFI-400437 (CAS#1247000-76-5, Cat# SYN-1207) (SynKinase, San Diego, CA, USA) [60 (link)]; centrinone (CAS#1798871-30-3, Ludwig Institute for Cancer Research, New York, NY, USA) [59 (link)]; centrinone B (CAS#1798871-31-4, Ludwig Institute for Cancer Research, New York, NY, USA) [59 (link)]; KW-2449 (CAS# 1000669-72-6, Cat#HY-10339) (MedChemExpress, Monmouth Junction, NJ, USA) [63 (link)]; R1530 (CAS#882531-87-5, Cat#15225) (Cayman Chemical, Ann Arbor, MI, USA) [57 (link)]; axitinib (CAS#319460-85-0, Cat#13813) (Cayman Chemical, Ann Arbor, MI, USA) [61 (link)], and alisertib (CAS#1028486-01-2, Cat# HY-10971) (MedChemExpress, Monmouth Junction, NJ, USA) [97 (link)] (Figure 1).