Synergy 2 microplate reader

Cells were seeded onto a 96-well plate and cultured in a standard culture medium for 24 h. Then, cells were washed in PBS, and the medium was changed to experimental (DMEM with 0.2% BSA), supplemented with different concentrations of ostarine, and cultured for the next 24 h. Some cells were grown in standard medium as a positive control.
After 24 h, to investigate viability, cells were treated with MTT solution ((3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) for 1 h. Then, the medium was removed, and 100 μL of DMSO was added to the cells to dissolve the formazan crystals. The optical density was measured using a Synergy 2 microplate reader (BioTek Instruments, Winooski, VT, USA).
Cell Proliferation ELISA BrdU colorimetric kit (Roche Diagnostics, Penzberg, Germany) was used to test proliferation by evaluating the incorporation of bromodeoxyuridine (BrdU) into new DNA strands of proliferating cells. After 21 h incubation with the examined concentrations of ostarine, BrdU solution (10 μM) was added for 3 h. The assay was performed according to the manufacturer’s instructions. The optical density of these samples was also measured using a Synergy 2 microplate reader (BioTek Instruments, Winooski, VT, USA).

Cells were seeded in 96-well clear, flat-bottom plates (Microtest 96, Falcon, Corning, NY, USA) at a density of 500 cells per well. Each plate was incubated overnight at 37 °C under 5% CO₂. R1881 and bicalutamide were dissolved in DMSO and diluted to the appropriate concentrations to a total volume of 100 μL and 0.1% DMSO per well. The cells were then incubated for at 37 °C. Cell viability was assessed by Celltiter-Blue assay kit (Promega, Madison, WI, USA) as instructed by the manufacturer. After treatment, cells were incubated with CellTiter-Blue^® reagent for 1 h at 37 °C. Fluorescence was measured on a BioTek Synergy 2 microplate reader (BioTek, Winooski, VT, USA) at 560/590 nM. For sulforhodamine B (SRB) assays, MOE cells were plated at 5 × 10³ cells/mL in a 96-well plate then treated and incubated for 5 days followed by colorimetric assay as described previously [35 (link)]. Absorbance at 505 nM was measured on a BioTek Synergy 2 microplate reader.

The NPs’ loading capacity
(LC) and encapsulation efficiency
(EE) were obtained indirectly by quantifying the unloaded drug in
the supernatant.
Free TMZ was quantified by fluorescence at
excitation and emission wavelengths of 420 and 540 nm, respectively
(Synergy 2 microplate reader, BioTek, UK). UV–vis absorbance
measurements quantified free BTZ at λ_max = 269 nm
(Synergy 2 microplate reader, BioTek, U.K.). The obtained absorbance
and fluorescence values were correlated to the BTZ and TMZ calibration
curves in PVA, respectively.
The EE values were expressed as
the percentage of the entrapped
drug to the total used drug. The LC of the NPs was given as the percentage
of the entrapped drugs to the PLGA weight.