Edu 5 ethynyl 2 deoxyuridine labeling solution

100 μL 8000 MM.1S, NCI-H929 cells of each group per well were cultured in 96-well plates for 24 h and then were cultured for 24, 48 and 72 h. At each time point, the CCK-8 assay (Solarbio, Beijing, China) was used to evaluate cell viability upon cells according to the manufacturer’s protocol. Cell viabilities were calculated by measuring the optical density at 450 nm, using a spectrophotometric plate reader (BioTek, VT, USA). All cell viability results were tested by three independent experiments.
200 μL 2 × 10⁴/mL cultured MM cells were fixed with 70% alcohol and then incubated with 50 μM EdU (5-Ethynyl-2′-Deoxyuridine) labeling solution (Invitrogen) at 37 °C for 2 h. According to the Click-iT™ EdU imaging kit (Invitrogen), the fluorescent intensity of EdU was determined at 550 nm. For subsequent DNA staining, cells were incubated with 5 μg/mL Hoechst 33342 for 30 min. Immunostainings were visualized and photographed with a fluorescent microscope (Olympus inverted microscope IX71) and calculated by flow cytometry.

1 × 10³ CC cells were maintained in 96-pore plates for 1 d and grown for extra 0, 24, 48, 72 and 96 h. Cell Counting Kit-8 (Bosterbio, Wuhan, China) was utilized to evaluate cell viability following the supplier’s protocol at each time point. After 4 h of culture, a spectrophotometric plate reader (BioTek, VT, USA) was applied to measure the optical density at 450 nm for detection of cell viability.
CC cells (200 μL, 2 × 10⁴/mL) were immobilized by 70% alcohol and subsequently cultured with 50 μM EdU (5-Ethynyl-2′-Deoxyuridine) labeling solution (Invitrogen) for 2 h at 37 °C. The fluorescent intensity of EdU was examined at 550 nm through applying Cell Light EdU DNA imaging kit (Invitrogen). Cells were cultivated using 5 μg/mL Hoechst 33342 for 0.5 h for DNA staining. The visualization and photograph of immunostainings were implemented with a fluorescent microscope (Olympus inverted microscope IX71).