Fig. 2: The loss of ezrin impacts the number of lung monocyte-derived IMs in response to LPS.

WT and Ezr-KO^m mice were nebulized with LPS (12.5 mg/5 ml for 15 min). The mice were sacrificed post-24 h of LPS and collected blood, BALF and lung tissue samples for flow cytometry and RNA analysis. A Numbers and percentages of lung moMs and IMs in lung tissue homogenates (inferior lobe) were quantified by sequential gating strategy from flow cytometry. Cell numbers were calculated from the percentage of viable cells multiplied by the total cell count in the inferior lung lobe. The gating strategy is described in Fig. S2C. B Flow cytometry-based dot plot representation of moMs, transitional IMs (tnsIMs) and mature IMs (matIMs) populations in WT, Ezr-KO^m and CCR2-KO mice, untreated or treated with LPS. C Numbers and percentages of moMs, tnsIMs and matIMs (quantification of (C)) in the lung (inferior lobe) tissue, untreated or treated with LPS. Quantification was performed as in (B). D Number (left) and percentage (right) of monocytes in the peripheral blood. Data are represented as mean ± SEM from three independent experiments with three or more mice per genotype. Each dot represents a biological replicate. Statistical analysis was performed using one-way ANOVA or Tukey’s test for multiple comparisons between the genotype and treatment conditions. *p < 0.05, **p < 0.01 and ns non significant. See also Supplementary Figs. S3–5.