Figure 2. Effects of ectopic miR-146a expression or miR-146a silencing on the Ly-6C^hi monocyte response.

a) Monocyte counts in spleen of mice reconstituted with miR-146a–expressing (miR-146a^hi) or control (miR-146a^norm) vector and challenged with Lm (n=3–5 from 2 independent experiments).

b) Number of TNFα-producing monocytes after ex vivo re-stimulation (same mice as in a).

c) Monocyte counts in spleen of mice that received either anti-miR-146a LNA or PBS and were challenged with live Lm for 24 h (n=3).

d) Number of TNFα-producing monocytes after LNA treatment (same mice as in c).

e) Ly-6C^hi/Ly-6C^lo monocyte ratios in wild-type (1.03±0.03) and miR-146a^−/− (1.05±0.04) mice in steady-state (mean±SEM).

f) Fold increase of Ly-6C^hi and Ly-6C^lo miR-146a^−/− monocytes in peritoneal cavity compared to their wild-type counterparts in bone marrow chimeras 4 d after peritoneal LPS injection (n=4 from 2 independent experiments).

g) Number of TNFα-producing wild-type or miR-146a^−/− monocytes (same mice as in f).

h) Colony forming unit (CFU) assay to quantify viable Lm from the spleen of wild-type (n=6) and miR-146a^−/− (n=5) mice 24 h after infection.

i) Fold increase of Ly-6C^hi and Ly-6C^lo miR-146a^−/− monocytes compared to their wild-type counterparts in bone marrow chimeras 24 h after peritoneal thioglycollate injection (n=3).

j) Tracking of EGFP⁺ wild-type and EGFP⁻ miR-146a^−/− CD45.2 GMP progeny. Right dot plot shows CD45.2 Lin⁻ CD11b⁺ CD115⁺ donor GMP-derived monocyte (representative of 4 independent experiments).

k) Fold increase of miR-146a^−/− neutrophils and monocytes (GMP donor-derived) compared to their wild-type counterparts in the peritoneal cavity 4 d after LPS challenge (n=4 from 2 independent experiments).

l) Ly-6C expression by donor GMP-derived monocytes (same mice as in k).

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