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. 2013 Dec 23;111(1):E109–E118. doi: 10.1073/pnas.1312585111

Fig. 1.

Fig. 1.

FRCs and endothelial cells expand rapidly during immune response in draining peripheral LNs. (A–D) Splenocytes from OT-1 (OVA-specific CD8+ TCR tg) and OT-2 (OVA-specific CD4+ TCR tg) C57BL/6 mice on a CD45.1+ background were transferred into C57BL/6 (CD45.2+) recipient mice, which then received s.c. injections of OVA/Mont or PBS, along with BrdU administration. The six draining peripheral LNs were isolated at the indicated time points after immunization, digested, and then analyzed by flow cytometry. (A) Number (Upper) and proliferation (BrdU incorporation; Lower) of indicated cell populations from PBS-control (open circles) and OVA/Mont-immunized (closed circles) mice. (B) Representative dot plots of DAPI−CD35−CD45− pregated stromal cells identifying FRCs (gp38+CD31−), LECs (gp38+CD31+), and BECs (gp38−CD31+). Percentages indicated show the cell frequency among all living cells. (C) Representative histograms showing FRCs stained with an antibody to BrdU (white) or an isotype-matched control (gray area) at 8.5 d after PBS or OVA/Mont injection. (D) Number (Upper) and proliferation (Lower) of FRCs, LECs, and BECs at indicated time points after OVA/Mont immunization. (E) WT C57BL/6 mice were infected with L. major in both footpads. On day 19, the two draining popliteal LNs were isolated, enzymatically digested, and analyzed as described above. Data are mean ± SD, representative of two or three independent experiments with n ≥3 mice per group.