The cells of each Petri dishes of control and experimental groups were maintained in MEM and RPMI 1640 media, where as it is applicable, supplemented with 10%% FBS, 50 μg/ml gentamycin, 50 μg/ml penicillin, and 50 μg/ml streptomycin at 37 °C in a 95%air/5%5 % CO2 atmosphere in CO2 incubator.
The cells of each Petri dish of control and experimental groups were maintained in MEM and RPMI 1640 media, where it is applicable, supplemented with 10%% FBS, 50 μg/ml gentamycin, 50 μg/ml penicillin, and 50 μg/ml streptomycin at 37 °C in a 95%air/5%5 % CO2 atmosphere in CO2 incubator.
Transfected cells were seeded in 100 mm-diameter dishes at variable densities and, when applicable, supplemented with cells transfected with the replication-deficient RNA neo-GAA to reach a final density of 5×105 cells per dish.
Typically 0.75 µg of ISG15variant, if indicated supplemented with 0.25 µg E1 and 0.5 µg E2 or, applicable supplemented with 0.75 µg substrate was used in a total volume of 250 µL DNA/CaPO4-transfection mixture.
Owing to their similarities in presentation, there is a tendency for government authorities to consider that all requirements for medicines are equally applicable to supplements, and this includes the issue of stability testing.
Where applicable, media were supplemented with 5 mM creatine, 50 mM glucose or 2.5 ng/mL TGF-β (Sigma-Aldrich).
If applicable, cultures were supplemented with L- tosylamido-2-phenyl)ethyL- tosylamido-2-phenyltrypsin at a finaL- tosylamido-2-phenyl µg/mL- tosylamido-2-phenyl
Cyanobacterial and L-BMAA hypotheses are tested via questions about: (1) drinking water; (2) bathing habits; (3) food consumption including dietary supplements (to be specified if applicable), the type of supplement is informed and (4) irrigation water, if any.
To this end, it is desirable to develop cost-effective and easily applicable age tracers/techniques to supplement the existing ones.
Where applicable, the medium was supplemented with PD98059 (MEK1-inhibitor) at the indicated concentration.
