Figure 3. Altered proliferation and trafficking of miR-146a^−/− Ly-6C^hi monocytes during inflammation.

a) Cytokine production of sorted wild-type and miR-146a^−/− monocyte subsets after in vitro LPS stimulation. Cytokine production is expressed per cell (n=2–3).

b) Time course TNFα production by Ly-6C^hi monocytes upon LPS challenge in vitro (n=2).

c) Gating strategy for DAPI staining of bone marrow Ly-6C^hi monocytes. Histograms show data for LPS stimulated wild-type or miR-146a^−/− animals.

d) Quantification of cell cycle status in wild-type and miR-146a^−/− animals in steady state (n=2) or after 4 consecutive days of LPS injection i.p. (n=4) in bone marrow, spleen and peritoneal cavity.

e) Number of donor wild-type and miR-146a^−/− EGFP⁺ CD45.2 Ly-6C^hi monocytes retrieved in the peritoneal cavity 6 h after transfer into LPS-treated CD45.1 recipient mice (n=4).

f) Flow cytometry-based cell surface CCR2 mean-fluorescence intensity (MFI) in wild-type and miR-146a^−/− blood monocytes (n=8).

g) In vitro chemotactic activity of wild-type (EGFP⁺) and miR-146a^−/− (EGFP⁻) Ly-6C^hi monocytes toward MCP-1 (n=4).

Data are presented as mean±SEM. (* p<0.05, ** p<0.01, *** p<0.001, Student’s t-test).