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. Author manuscript; available in PMC: 2015 May 12.
Published in final edited form as: Nat Protoc. 2014 Jan 2;9(1):209–222. doi: 10.1038/nprot.2014.005

Figure 1.

Figure 1

Evidence that perfusion should be avoided. (a) Anti-CD45.2 mAb was injected i.v. via the tail vein of naive C57BL/6 mice 3 min before perfusion and lymphocyte isolation. Naive CD4 + and CD8 + T cells and B cells were identified by flow cytometry, as indicated and examined for labeling with injected anti-CD45.2 mAb. Representative of 12 mice from two experiments. (b) At 15 d after i.t. LCMV infection, lungs from P14 chimeras were perfused or left unperfused. Representative immunofluorescence images of iBALT or aggregate structures in lungs stained with anti-collagen type IV (green), anti-PNAd (red), anti-Thy1.1 (gray) and DAPI (blue). Arrow designates PNAd + high endothelial venule (HEV). Scale bars, 100 µm. Representative of two experiments totaling 10 mice per condition. *P = 0.05, **P < 0.01, using a two-tailed Student’s t test with a 95% confidence interval. Error bars indicate s.e.m.