Raw data for measured competitive ELISA and cellular assays for HVEM/LIGHT complex inhibition using peptides

<p>The research system was the HVEM/LIGHT complex, the inhibition of which was measured using peptides. An ELISA was performed using 96-well plates coated with LIGHT-His protein, followed by blocking with BSA. Peptides at three concentrations were added, then HVEM-Fc and an HRP-conjugated anti-human IgG antibody. After incubation and washing steps, TMB was added, and absorbance was measured at 492 and 650 nm. Wells without peptides served as controls. Cell-based assays were conducted using J-NF-κB::eGFP HVEM and TCS LIGHT cell lines maintained in RPMI 1640 with antibiotics and 10% heat-inactivated FBS. Peptides at three concentrations were preincubated with TCS LIGHT cells, washed, and transferred to 96-well plates. J-NF-κB::eGFP HVEM cells were then added, and the plate was incubated for 24 h. After staining with APC-conjugated mCD45 antibody to exclude TCS cells, eGFP expression was measured by flow cytometry. </p>