Celltiter 96 s aqueous one solution cell proliferation assay mts

Zirconium tetrachloride (ZrCl 4 , 99.5%), terephthalic acid (BDC, 98%) and naphthalene-2,6-dicarboxylic acid were bought from Alfa Aesar (UK). Benzoic acid (99.5%), HCl (37%), dimethylformamide (DMF, 99.8%), L-proline, 2-aminoterephthalic acid, methanol (99.9%), acetone (99.9%), calcein disodium salt and a-cyano-4-hydroxycinnamic acid (a-CHC) were obtained from Sigma-Aldrich (UK). 2-Bromoterephthalic acid and 2-nitroterephthalic acid were obtained from Acros Organics. Azobenzene-4,4 0 -dicarboxylic acid and 4,4 0 -stilbenedicarboxylic acid were bought from TCI UK. 4,4 0 -Biphenyldicarboxylic acid was obtained from Fluorochem.
HeLa cells were obtained from the ATCC. Dulbecco's modified Eagle's medium (DMEM), foetal bovine serum (FBS), L-glutamine, penicillin, and streptomycin were purchased from Invitrogen (UK). Phosphate-Buffered Saline (PBS) and trypsin-EDTA, were purchased from Life Technologiest (UK). The CellTiter 96 s Aqueous One Solution Cell Proliferation Assay (MTS) was obtained from Promega (UK). All chemicals and biochemicals used were of analytical grade.

All solvents were of analytical or HPLC grade and purchased from Sigma or Fisher Scientific unless otherwise stated. Deuterated solvents were from Sigma. Acryloyl chloride, ethylene-dioxy-bisethylamine, triethylamine (TEA), dithiodiethanol, sodium chloride, potassium phosphate dibasic and potassium phosphate monobasic, sodium azide, sodium phosphate dibasic, ethidium bromide (EtBr), fluorescamine, polyethylenimine (PEI) (25 kDa, branched) calf thymus DNA and glutathione (GSH), were used as received from Sigma Aldrich. Luciferase siRNA (CCGCAAGAUCCGCGAGAUU) was provided by Eurogentec (UK). Luciferase Assay System with Reporter Lysis Buffer and CellTiter 96 s AQueous One Solution Cell Proliferation Assay (MTS) were provided by Promega (UK). Silencert Cyt3-labeled Negative Control was provided by Thermofisher Scientific (UK).

A549 luciferase expressing cells (2 Â 10 4 ) were placed in 96-well plates and cultured in 200 mL of cell medium with or without FBS at 10%. After 24 h, cells were treated with free OBAEs in the concentration range 0.005-5 mg mL À1 . As control, cells were treated as well as with an aqueous solution of free polyethylenimine (PEI) at the same concentration range. Cells treated with 0.1% (v/v) Triton-X 100 and fresh media were used as a positive and a negative control, respectively. After 24 h of incubation, cells were washed with PBS and treated with CellTiter 96 s Aqueous One Solution Cell Proliferation Assay (MTS, Promega) (20 mL per well). After further incubation (3 h), the absorbance was read at 490 nm in a microplate reader (Tecan Platereader). The percentage of metabolic activity (%) was calculated according to the equation: Cell viability (%) = [(OD sample À OD CTR+)/ (OD CTR À OD CTR+)] Â 100