CIN was induced in RPE TP53−/− cells using a modified protocol described previously51 . In brief, the cells were synchronized at the G1/S border with 5 mM thymidine (Sigma-Aldrich, T9250) for 24 h. Six hours after thymidine block release, the cells were treated with 500 nM of the MPS1 inhibitor reversine52 (link) (Sigma-Aldrich, R3904) for 12 h. Before processing for downstream analyses, cells were allowed to recover for 6 h.
R3904
Manufactured by Merck Group
Sourced in United States
R3904 is a laboratory equipment product manufactured by Merck Group. It is designed for general laboratory use, providing a basic functionality for various scientific applications. The core function of R3904 is to facilitate standard laboratory procedures and tasks. No further details or interpretations are provided.
Automatically generated - may contain errors
2 protocols using r3904
1
Inducing CIN in TP53-/- RPE Cells
2
Reversine Cytotoxicity Assay in Cell Lines
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MNNG/HOS, U-2 OS and MG-63 cell viability were determined using CCK8 assays (Dojindo Molecular Technologies, Japan). All cells were cultured in 96-well plates (2.5 × 104 cells/mL) and treated with different concentrations of reversine (1, 2, and 4 μM; Sigma-Aldrich, R3904, St. Louis, MO, USA). After 24 and 48 h culture, 10 μL of CCK8 (5 g/L) was added into the 96-well plates, followed by further incubation for 4 h at 37 °C. The CCK8 reactions were measured in Bio-Rad Model 450 Microplate Reader (Bio-Rad, Hercules, CA, USA) with 450 nm absorbance. Cell viability was indirectly determined using the ratio of the absorbance value for reversine-treated cells relative to the control cells (n = 6 for each group).
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