Brdu incorporation assay kit

Proliferation of the transfected cells plated in triplicate wells was determined by a BrdU incorporation assay kit (Roche Diagnostics) as per the manufacturer's protocol [4 (link)]. Data are presented as mean ± S.E. of duplicate experiments.

Cells (1 × 10⁴) per well were seeded in a 96-well plate and treated with miRNA for 48–72 h. Cell viability was evaluated using a cell counting kit-8 (CCK-8; Dojindo, Kumamoto) [73 (link)]. After incubation with CCK-8 solution for 1–2 h, a Synergy HTX (BioTEK, Winooski, VT) multi-mode reader was used for evaluating cell viability by measuring optical density (OD) at 450 nm [73 (link)]. A bromodeoxyuridine (BrdU) incorporation assay kit (Roche Diagnostics, GmbH, Mannheim, Germany) was employed for evaluating cell proliferation rate according to manufacturer’s protocol [72 (link)].

Cell proliferation assay was performed with BrdU incorporation assay kit according to manufacturer’s protocol (Roche Diagnostics). Briefly, normal and glaucomatous TM cells were transfected with pEGFP-Vector, pGFP-Prdx6 were seeded into 96-well plates at a density of 5×10³ cells per well. Following incubation cells were labeled with BrdU for 24 h and O.D. was measured at 450 nm (DTX 880 Multimode Detector, Molecular Device).

Cell proliferation was evaluated using a BrdU incorporation assay kit (Roche, Indianapolis, IN, USA) as previously described [59 (link)]. Briefly, HaCaT cells were plated in a 96-well plate with 2 × 10³ cells per well and were treated with various concentrations of CSFAb dissolved in DMEM containing 0.5% DMSO or a positive control EGF (50 ng/mL) for 36 h. BrdU labeling solution (10 μM) was then added and incubated at 37 °C for 12 h. The cells were then fixed, to denature DNA, with FixDenat solution from the BrdU kit for 30 min at RT, followed by incubation with peroxidase-labeled anti-BrdU monoclonal antibody at RT for 90 min. BrdU antibody complexes were detected using a luminometer (Synergy 2; Bio-Tek Instruments). Cell proliferation levels were expressed as percentages of those of untreated controls.

OT-I CD8⁺ T cells were cocultured with DCs that had been exposed to OVA protein plus PC nanogel or alum for 3 days, as described above, and cell proliferation was examined with a BrdU incorporation assay kit (Roche).

Cells (5 × 10³ cells/well) were seeded in a 96-well plate. The cell counting kit-8 (CCK-8; Dojindo, Kumamoto, Japan) assay was utilized to determine cell viability. CCK-8 solution was added to each well and the plates were incubated for 1–2 h. The optical density was quantified at 450 nm using a microplate reader Synergy HTX (BioTek Instruments, Inc., Winooski, VT, USA). Cell viability was presented as a fold change from the control.
A bromodeoxyuridine (BrdU) incorporation assay kit (Roche Diagnostics, GmbH, Mannheim, Germany) was used for the determination of cell proliferation rate. Cells were seeded at 5 × 10³ cells/well in 96-well plates and incubated for 48 h at 37 °C and 5% CO₂. BrdU was added to each well and the plates were incubated at 37 °C and 5% CO₂. The labeling medium was removed and a FixDenat solution was added to each well. After incubation for 30 min, FixDenat solution was removed. Anti-BrdU-POD solution was added to each well and incubated for 90 min at room temperature. The cells were then washed with PBS for three times and incubated with substrate solution for 20 min at room temperature. Then, 2 N H₂SO₄ was used as a stopping solution. The absorbance was quantified at 450 nm using a microplate reader Synergy HTX. Proliferation rates were presented as the fold change from controls.