Trustainfcx anti mouse cd16 32 fcr block

Cells were resuspended in 100 ml FCB, blocked with 0.5 ml TruStainFcX (anti-mouse CD16/32 FcR block, Biolegend) for 5 min at room temperature, and stained for 30 min at 4°C with: FITC-anti-CD45 (1:1000, Biolegend), PE-Cy7-anti-CD4 (1:200, eBioscience), PerCP-Cy5.5-anti-CD8 (1:200, Biolegend), PE-anti-B220 (1:200, Biolegend), APC-anti-MHCII (1:200, eBioscience), and APC-eFluor780-anti-CD11b (1:200, eBioscience) (Supplemental Table 1). Cells were then xed in 1% PFA for 1 h. Finally, cells were resuspended in 500 ml FCB, labelled with DAPI to exclude cell debris and analyzed with a CytoFLEX S ow cytometer (Beckman-Coulter) equipped with CytExpert software (Beckman-Coulter).
Quanti cation of IC100 in Tissues. IC100 was quanti ed in brain, spinal cord, liver and spleen at 35 days post-induction (dpi) of EAE using a proprietary assay developed by In amaCORE, LLC using Meso Scale Technology. Protein lysates were obtained as described in [29] . The assay was read using the QuickPlex SQ 120 instrument (Meso Scale Diagnostics, Maryland).