Epr6855

Formalin-fixed, paraffin-embedded (FFPE) skin biopsy tissue sections from lesional skin of patients with ACLE, SCLE, or DLE were analyzed using the Hyperion imaging CyTOF system (Fluidigm) as previously described (24 (link)) with modifications of the antibody panel. Specifically, metal-tagged antibodies including pan-keratin (C11, Biolegend), BDCA2 (Polyclonal, R&D Systems), CD56 (123C3, ThermoFisher Scientific), HLA-DR (LN3, Biolegend), CD11c (EP1347Y, Abcam), and CD4 (EPR6855, Fluidigm) were added in this study.

Tissue sections of 10 μm thickness from the liver and spleen of iMCD-NOS NSG2 mice were processed and stained according to the protocol of the Hyperion Imaging System provided by Fluidigm. The regions of interest detected by hematoxylin-eosin staining of slides of the liver and spleen were chosen for analysis. The protein expression levels of cells residing within the corresponding regions were quantified and visualized using a Hyperion Imaging System (Fluidigm). The following antibodies were used: anti-human CD4 (EPR6855, cat# 3156033D, Fluidigm, ¹⁵⁶Gd, 1:100), anti-human CD8a (C8/144B, cat# 3162034D, Fluidigm, ¹⁶²Dy, 1:50) and anti-human CD20 (H1, cat# 3161029D, Fluidigm, ¹⁶¹Dy, 1:200). Images of slides were prepared from PDX mice with iMCD NSG2. We obtained data of IMC from at least three independent experiments. In brief, IMC imaging mcd.files were exported into tiff files using the R software and MCD viewer (Fluidigm).