Abstract

<p><b>A</b>: Schematic representation for the isolation of specific splenic cell types from mice. Splenocytes were released by repeated collagenase digestion from freshly dissected spleens, followed by removal of erythrocytes and purification of splenocytes on Lympholyte M gradients. Splenic cell types are isolated by positive selection with magnetic beads coated with cell type-specific mAbs as specified. <b>B</b>: The purities of MACS-isolated cells were analysed by FACS using cell-type specific mAbs and isotype controls as specified in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1002538#s4" target="_blank">Materials and Methods</a>. One representative out of three experiments is shown. (Bv) CD11<sup>low</sup> B220<sup>+</sup> pDCs, isolated with murine plasmacytoid dendritic antigen-1 (mPDCA-1) showed a purity of about 90% in three independent experiments. (Bvi) The macrophage population, isolated with CD11b microbeads after depletion of CD11c<sup>+</sup> cells was contaminated with CD11c<sup>+</sup> CD11b<sup>+</sup> mDCs. Macrophages were therefore isolated by FACS instead (<b>C</b>). <b>C</b>: Splenocytes labelled with mAbs against anti-CD11b (M1/70) and anti-CD11c (HL3) were isolated by FACS using a DAKO cell sorter.</p

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