In vivo cytokine assessment

Anesthetized mice were bled and 50 to 100 μL of serum harvested. For experiments requiring larger volumes, mice were euthanized followed by terminal bleeding. Serum was frozen in −80°C, then thawed once for cytokine assessment. Mouse cytokine array Q1 (RayBiotech, Norcross, GA) was used according to the manufacturer’s instructions. Plates were read using GenePix 4000B (Molecular Devices). For confirmation, serum was evaluated with mouse IFN-γ Quantikine ELISA and mouse IL-6 Quantikine ELISA (R&D systems), as previously described.