The proliferation ability was determined by the EdU proliferation assay (RiboBio). EdU is a thymidine analogue, and its attached alkyne group is rare in natural compounds. It can be incorporated as thymine (T) into the DNA molecule being synthesized during DNA replication. Based on the specific reaction of Apollo® fluorescent dyes with EdU, DNA replication activity can be directly and accurately detected. After different treatments of HCT116‐lohp and sw480‐lohp cells (etomoxir 150 μm , oxaliplatin 2 μg·mL−1. siRNA/NC or combined according to the experiment), all steps were carried out in accordance with the instructions of the EdU proliferation assay (RiboBio). EdU (50 μm ) was added to the cells, which were then incubated for 5 h, fixed, filtered and stained according to the instructions. Finally, the results were imaged by a Leica microscope (blue represents all cells, red represents proliferating cells).
Edu proliferation assay
Manufactured by RiboBio
Sourced in China
The EdU proliferation assay is a laboratory tool used to measure cell proliferation. It detects the incorporation of the nucleoside analog EdU (5-ethynyl-2'-deoxyuridine) into the DNA of dividing cells, allowing for the quantification of cell proliferation.
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Lab products found in correlation
Cell counting kit 8 cck 8 assay, by Beyotime (2 mentions)
Hoechst 33342, by RiboBio (2 mentions)
Apollo staining solution, by RiboBio (2 mentions)
Fluorescence microscope, by Olympus (2 mentions)
Uorescence microscope, by Leica (2 mentions)
Edu assay, by RiboBio (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Dmi6000b inverted microscope, by Leica (1 mentions)
Fluorescence microscope, by Leica (1 mentions)
Infinite m200 multimode microplate reader, by Tecan (1 mentions)
Cck 8, by Dojindo Laboratories (1 mentions)
Edu assay kit, by RiboBio (1 mentions)
Dmi4000b, by Leica (1 mentions)
Spectramax m5 multi mode microplate reader, by Molecular Devices (1 mentions)
Cck 8, by Yeasen (1 mentions)
Fv300, by Olympus (1 mentions)
Fluorescence microscope, by Nikon (1 mentions)
Cck 8 assay, by Dojindo Laboratories (1 mentions)
29 protocols using edu proliferation assay
1
Evaluating Cell Proliferation with EdU Assay
2
Evaluating Cell Proliferation with EdU Assay
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To display the function of miR‐505 on cell proliferative, the EdU proliferation assay (RiboBio, Nanjing, China) was carried out according to the manufacturer's instructions. Twenty‐four hours after transfection, cells were incubated with 50 μM EdU for 2 hr. Then an Apollo staining and 4′,6‐diamidino‐2‐phenylindole (DAPI) staining were performed according to the instructions to detect the EdU‐positive cells (red cells) with a fluorescence microscope. The EdU incorporation rate was revealed as the ratio of EdU‐positive to total DAPI‐positive cells (blue cells).
3
Cell Proliferation Assays for Exendin-4
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Cell proliferation was assessed with the cell counting kit-8 (CCK-8) assay (Beyotime Institute of Biotechnology, China) and 5-ethynyl-2´-deoxyuridine (EdU) proliferation assay (RiboBio Co., China). For the CCK-8 test, cells were plated onto 96-well plates (5 × 103 cells/well) with Ex-4 (0–20 nm/L) in a triplicate pattern. Assays were performed from 1 to 7 days after plating by the addition of 100 μl of fresh medium in 10 μl of the CCK-8 solution for another 2 h at 37 °C. The OD at 570 nm was measured. The assay was repeated 3 times.
The EdU assay (RIBOBio Co, Guangzhou, China) was used to measure cells’ ability to proliferate after treatment with 20 nm Ex-4 for 24 h. After incubation with EdU for 2 h, the cells were fixed with 4% paraformaldehyde and permeabilized with 0.5% Triton X-100. Then, the Apollo® reaction cocktail (reaction buffer and Apollo® 643 fluorescence) was added to the medium for another 30 min in the dark. After being washed with PBS 3 times, the cells were stained with DAPI (Sigma-Aldrich) for 5 min and immediately viewed under fluorescence microscopy. The number of EdU+ cells was calculated by counting at least three random separate fields.
The EdU assay (RIBOBio Co, Guangzhou, China) was used to measure cells’ ability to proliferate after treatment with 20 nm Ex-4 for 24 h. After incubation with EdU for 2 h, the cells were fixed with 4% paraformaldehyde and permeabilized with 0.5% Triton X-100. Then, the Apollo® reaction cocktail (reaction buffer and Apollo® 643 fluorescence) was added to the medium for another 30 min in the dark. After being washed with PBS 3 times, the cells were stained with DAPI (Sigma-Aldrich) for 5 min and immediately viewed under fluorescence microscopy. The number of EdU+ cells was calculated by counting at least three random separate fields.
4
Caco2 Cell Proliferation Assay
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To perform the cell proliferation, EdU proliferation assay (RiboBio, Nanjing, China) was utilized. Caco2 cells were treated with 50 μM EdU for 2 hours. After the incubation, cells were fixed and stained by EdU. A Leica DMI6000 B inverted microscope was used to take the images.
5
Measuring miR-182-5p Impact on Cell Proliferation
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To investigate the influence of miR-182-5p on cell proliferation, EdU proliferation assay (RiboBio, Guangzhou, China) was conducted. Briefly, cells were incubated with 50 μM EdU for 2 h at 37 °C. Cells were fixed with 4% paraformaldehyde and treated with 0.5% Triton X-100 at room temperature. Next, the cells were washed with phosphate buffered saline (PBS) and incubated with Hoechst 33342 (100 μL) at room temperature for 30 min. The EdU positive cells were then visualized under a fluorescence microscope (Leica, Germany).
6
Evaluating microRNA-15a's Impact on Cell Proliferation
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To detect the function of microRNA-15a on cell proliferation, 5-ethynyl-2′-deoxyuridine (EdU) proliferation assay (RiboBio, Nanjing, China) was conducted. After transfection for 48 h, cells were incubated with 50 μM EdU. An Apollo and DAPI staining were employed to detect the EdU-positive cells.
7
Cell Proliferation Assays: CCK8 and EdU
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CCK8 (Dojindo, Japan) assay was performed. The cells were cultivated on a 96-well plate (5.0 × 103 cells per well) for 24 h after transfection. TECAN infinite M200 Multimode microplate reader (Tecan, Mechelen, Belgium) was then employed after an hour of incubation in CCK8 at the absorbance of 450 nm.
EdU proliferation assay (RiboBio, Nanjing, China) was conducted to detect the proliferation of the transfected cell. The cells were fixed by 4% paraformaldehyde for half an hour after being treated with 50 μM EdU for 2 h. Following Apollo staining and DAPI staining, a fluorescent microscope was adopted to observe the EdU positive cells.
EdU proliferation assay (RiboBio, Nanjing, China) was conducted to detect the proliferation of the transfected cell. The cells were fixed by 4% paraformaldehyde for half an hour after being treated with 50 μM EdU for 2 h. Following Apollo staining and DAPI staining, a fluorescent microscope was adopted to observe the EdU positive cells.
8
Omentin-1 Modulates Cell Proliferation
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Cell proliferation was assessed with the cell counting kit-8 (CCK-8) assay (Beyotime Institute of Biotechnology) and 5-ethynyl-2’-deoxyuridine (EdU) proliferation assay (RiboBio Co., China). For the CCK-8 test, cells were plated onto 96-well plates (3 × 103 cells/well) with omentin-1 (0–800 ng/ml; Omentin Human Recombinant; Prospect, Ness-Ziona, Israel) in a triplicate pattern. Assays were performed from 1 to 7 days after plating with the addition of 100 μl of fresh medium in 10 μl of CCK-8 solution for another 2 h at 37 °C. The optical density (OD) at 450 nm was measured. The assay was repeated three times.
For the EdU assay, cells were incubated with omentin-1 (800 ng/ml) for 5 days in 96-well plates, and then assessed using an EdU assay kit (RiboBio Co., China) according to the manufacturer's instructions. The EdU-positive cells were viewed under fluorescence microscopy (DMI4000B; Leica, Wetzlar, Germany) and the number calculated by counting at least three random separate fields.
For the EdU assay, cells were incubated with omentin-1 (800 ng/ml) for 5 days in 96-well plates, and then assessed using an EdU assay kit (RiboBio Co., China) according to the manufacturer's instructions. The EdU-positive cells were viewed under fluorescence microscopy (DMI4000B; Leica, Wetzlar, Germany) and the number calculated by counting at least three random separate fields.
9
Proliferation Assay of SGC7901 Cells
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The proliferative ability of SGC7901 cells after different transfection were determined by the EdU proliferation assay (RiboBio Inc.). Twenty-four hours after transfection, cells were incubated in 50 M EdU for 5 h, and fixed within 4% paraformaldehyde for 30 min at room temperature (RT). Then the cells were washed in PBS twice and permeabilized using PBS containing 0.5% Triton X-100 for 10 min. After extensive washes in PBS, the cells were incubated lucifugally in Apollo staining solution (RiboBio Inc.) for 30 min, then repeated permeation and wash, and incubated in Hoechst 33342 (1:100; RiboBio Inc.) for another 30 min at RT. All of the staining were performed in triplicate.
10
Proliferation Capacity of HPAEC Cells
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To assess the proliferative capacity of HPAEC cells, EdU proliferation assay (RiboBio, Guangzhou, China) was performed. After transfection for 24 h, the cells were incubated with 50 μm EDU for 2 h, then stained with AdoLo and 4′,6-diamidino-2-phenylindole (DAPI), and the number of EDU-positive cells was examined by fluorescence microscopy. The display rate of EDU positive was shown as the ratio of the number of EDU positive cells to the total DAPI chromogenic cells (blue cells).
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