Luminex Assay

The protein level of CXCL10 in the cell culture supernatant was assessed using the human Magnetic Luminex Assay (R&D Systems), which was performed per the manufacturer's instructions. Briefly, all the samples and standards were first mixed with the CXCL10 antibody coated magnetic microparticles and incubated for 2 h at room temperature on a horizontal orbital microplate shaker set at 800 rpm. After washing the microparticles, biotinylated detector antibodies were added and incubated for 1 h at room temperature on the shaker set at 800 rpm. Following a wash to remove any unbound biotinylated detector antibody, streptavidin-phycoerythrin conjugates were added and incubated for 30 min at room temperature on the shaker set at 800 rpm. Finally, the protein level of CXCL10 in the cell culture supernatant was analyzed using the Bio-Plex^TM 200 system (Bio-Rad).