Sunrise remote r

HAECs, HUVECs, or HASMCs at passages 2 to 8 were seeded into 96‐well plates (1×10⁴ cells/100 μL per well) and incubated for 24 hours in the same media (HuMedia‐EG2, EGM‐2, or SmGM‐2, respectively). Cells were then incubated for another 48 hours in fresh media with the indicated concentrations of neopterin in the absence or presence of anti–neopterin antibody (10 μL/mL) or specific inhibitors of c‐Src (0.1 μmol/L saracatinib) and MEK1 (MAPK/ERK kinase 1) or MEK2 (10 μmol/L SL327). Next, 10µL WST‐8 solution (Cell Count Reagent SF; Nacalai Tesque) was then added to each well.2, 3, 4, 5, 6, 7, 8 After 1 hour of incubation, the quantity of formazan product was determined by reading absorbance at 450 nm using a Sunrise Remote R microplate reader (Tecan).17, 18, 19, 20, 21, 22, 23

HUVECs, HUVEC‐derived EA.hy926 cells, or HASMCs at passage 2 to 8 were seeded into 96‐well plates (1×10⁴ cells/100 μL/well) and incubated for 24 hours in DMEM or SmGM‐2 containing 10% or 5% FBS, respectively. Cells were then incubated for a further 48 hours with the indicated concentrations of KP‐10 in fresh media. Ten microliters of WST‐8 solution (Cell Count Reagent SF; Nacalai Tesque, Inc, Kyoto, Japan) were then added to each well. After 1 hour of incubation, the quantity of formazan product was determined by reading absorbance at 450 nm using a Sunrise Remote R‐micro plate reader (Tecan, Männedorf, Switzerland).19, 20, 21, 22, 23