2mL of NGD was added to 100k pMGLs in a 35mm well and allowed to condition for 24 hours. For IFN-γ/LPS stimulation, LPS was added at 100ng/mL along with 20ng/mL IFN-γ at the onset. 400uL of this supernatant were added to a prepared cytokine antibody panel membrane, per the manufacturer’s instructions (RnD Systems, kits ARY005 and replacement ARY005B). After incubation, the membranes were revealed with ECL reagent.
