Cell light edu apollo 567 in vitro imaging kit

Cell proliferation was detected using a Cell Counting Kit-8 (CCK-8; Beyotime, Beijing, China) in accordance with the manufacture’s specification. GBM cells were plated into 96-well plates after transfection and counted every 24 h. The optical absorbance at 450 nm was measured. Cell viability was also determined with the 5-ethynyl-2′-deoxyuridine (EdU) assay using Cell-light EdU Apollo 567 In Vitro Imaging Kit (Ribobio, Guangzhou, China) following the manufacturer’s instructions. For the colony formation assay, cells were plated in 6-well plates and cultured in medium with 10% FBS to allow colony formation. The medium was replenished every 4 days. Two weeks later, cells were fixed with 96% ethanol and stained with 0.1% crystal violet. Colony formation ability was evaluated by counting the number of colonies consisting of more than 50 cells.

At 48 h after transfection, the 5-ethynyl-2′-deoxyuridine (EdU) incorporation assay was performed using the Cell-Light EdU Apollo567 In Vitro Imaging Kit (Ribobio) according to the manufacturer’s instructions. The xenograft tumor tissues were harvested in 4% formaldehyde buffered with phosphate-buffered saline, embedded in paraffin and then sectioned. An antibody against Ki-67 (#9449, Cell Signaling Technology, Danvers, MA, USA, dilution: 1/400) was used for immunohistochemical analyses. Immunoreactivity in the sections was detected using a horseradish peroxidase (3,3′-diaminobenzidine substrate) kit (BioGenex, Fremont, CA, USA). The slides were then counterstained with hematoxylin, dehydrated and mounted. Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL) (Roche) assays were carried out according to the manufacturer’s protocol.

First, HCT116 and SW480 cells were grown in a 5-ethynyl-2’-deoxyuridine (EdU) solution for 2 hr. The cells were fixed with PBS containing 4% paraformaldehyde. Finally, the fixed cells were deposited in 70% ethanol, and then Cell-Light™ EdU Apollo^®567 In Vitro Imaging Kit (RiboBio, China) was used to dye cells. Cell growth was observed by fluorescence microscopy.

Cell viability was measured with Cell Counting Kit-8 (Dojindo Laboratory, Japan) every 24 h. Cell proliferation ability was detected by the Cell-light™ EdU Apollo® 567 In Vitro Imaging Kit (Ribobio, Guangzhou, China) 48 h later. Cell cycle distribution was analyzed by flow cytometry (BD Biosciences, USA) using propidium iodide (PI) staining (Beyotime, Shanghai, China) after 48 h–transfection and overnight fixation.