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. 2011 Jun 27;6(6):e21341. doi: 10.1371/journal.pone.0021341

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Hou et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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A, Immunoprecipitation assay. The membrane protein extracted from the porcine 4Ig-B7-H3-transfected 293 T cells (Lane 1) or vacant plasmid-transfected 293 T cells (Lane 2) was pulled down with anti-porcine B7-H3 mAb-bound protein-G agarose. The pulled-down proteins were submitted to western-blot assay. Lane 3: anti-porcine B7-H3 mAb-bound protein-G agarose control. Lane 4: protein marker. B, The analysis of the porcine 2Ig-(left plot) and 4Ig-B7-H3 (right plot) expressed on 293 T cells stained by the anti-porcine B7-H3 mAb (filled histogram) or isotypic Ab (open histogram). C, non-activated monocytes were stained with a combination of anti-CD14 with either anti-B7-H3 mAb (filled histogram) or isotypic Ab (open histogram). D, The monocytes were co-cultured with IFN-γ in a final concentration of 30 ng/ml (left plots) or LPS at a final concentration of 1 µg/ml (right plots) for 24 h (upper plots) or 48 h plots (lower plots). On the indicated time points, the monocytes were harvested to be stained with a combination of anti-CD14 with either anti-B7-H3 mAb (filled histogram) or isotypic Ab (open histogram).