Figure 1.

Subcutaneous CpG injection mobilizes Ly6C^hi Monocytes and DCs. (A) C57Bl/6 mice (n = 4) were injected in the footpad with 20 µl of CpG (100 µg/ml) and blood samples and draining popliteal lymph nodes were analysed at the indicated time intervals by flow cytometry. (B,D) Flow cytometry plots showing the frequency of Ly6C^hi Monocytes in blood (B) and draining lymph node (D) of CpG injected mice at 0 h, 12 h and 96 h post CpG injection. (C,E) Graph showing the frequency of Ly6C^hi monocytes in blood (C) and draining lymph nodes (E) of CpG injected mice (as percentage of CD45+ Ly6G− leukocytes) over time. Mean values at each time interval where compared to mean values at steady state (0 h) ***P < 0.001, **p < 0.01, *p < 0.05. (F) Gating strategy applied to identify migratory DCs and resident DCs in popliteal lymph nodes. (G) Graph showing the absolute numbers of Ly6C^hi monocytes, migratory DCs and resident DCs in the draining lymph nodes of OVA/CpG injected lice at the indicated time intervals (n = 4; mean +/− SD). (H) Pie charts depicting the relative proportions of Ly6C^hi monocytes, migratory DCs and resident DCs in the draining lymph nodes. The area of the circles displayed is relative to the added counts of Ly6C^hi monocytes, migratory DCs and resident DCs at the indicated time intervals. Data shown are representative of three independent experiments.