Alexa fluor 594 beyoclick edu cell proliferation kit

The A498 and 786‐O cells (pretreated with si‐NC or si‐RCC1) and the 769‐P cells (transfected with OE‐NC/OE‐RCC1) were seeded in 96‐well plates, adding 2000 cells per well. Next, on appointed Day 0, 1, 2, 3, and 4, the cells were mixed with 10% CCK‐8 reagent (MCE) and incubated for 1 hour in the presence of 5% CO₂, at 37°C. Then, the value of OD 450 nm, indicating the relative cell viability, was measured using a Bio‐rad microplate reader.
For the colony formation assay, 1000 cells were plated into each well of 6‐well plates. After cultivation for 7–10 days, each well was washed using PBS (Procell) and stained with 4% paraformaldehyde and 0.5% crystal violet (Beyotime).
The EDU assay was also performed to detect cell viability using the Alexa Fluor 594 BeyoClick™ EdU Cell Proliferation Kit (Beyotime) according to the manufacturer's protocol.

The EdU assay was employed to measure the synthesis of new DNA and identify cell proliferation activity. Transfected cells (3 × 10⁵) in the logarithmic growth phase were inoculated into 6-well plates. The medium was changed after cell walling and co-cultured with V. parvula at MOI = 100 under anaerobic conditions for 12 h; the plates were then transferred to a 5% CO₂ incubator at 37 °C and incubated for 24 h. The culture medium was changed, followed by incubation with 10 μM EdU for 2 h, stained with Alexa Fluor 594 BeyoClick™ EdU Cell Proliferation Kit (Beyotime, C00788L), observed under a fluorescent inverted microscope (Vert. A1; Zeiss, Germany) and photographed.