Figure 8. Blocking G-CSF activity... | Rockefeller University Press

Figure 8.

$Figure 8. Blocking G-CSF activity induces neutrophil sequestration in the lungs, leading to sterile pulmonary inflammation in mice i.p. injected with E. coli. (A) Schematic representation of the experimental procedures. (B) Cells in BALF were stained with a modified Wright–Giemsa stain (40× immersion oil). (C) The percentage of neutrophils in BALF. (D) Neutrophil counts in BALF. (E) Staining of lung sections shows emigrated neutrophils in the lungs (20× dry lens). (F) The number of cells in the lung capillaries was quantified as the number of cells in total capillaries (but not alveolar air spaces)/10 alveolar air space area using ImageJ. (G) Emigrated neutrophils in alveolar spaces were quantified as volume fraction of the alveolar air space using standard point-counting morphometric techniques. (H) Cytokine/chemokine levels in BALF were measured using specific ELISA kits. (I and J) G-CSF blockage–induced elevation of cytokine levels in the lungs is mediated by neutrophils. (I) Schematic representation of the experimental procedures. Neutrophils were depleted by a Gr-1 antibody in both PB (∼85% depletion) and the BM (∼65% depletion; Kwak et al., 2015). (J) Cytokine/chemokine levels in BALF. Data shown are means ± SD of three experiments (n = 5 mice). *, P < 0.01 versus control (mice treated with IgG).$

Blocking G-CSF activity induces neutrophil sequestration in the lungs, leading to sterile pulmonary inflammation in mice i.p. injected with E. coli. (A) Schematic representation of the experimental procedures. (B) Cells in BALF were stained with a modified Wright–Giemsa stain (40× immersion oil). (C) The percentage of neutrophils in BALF. (D) Neutrophil counts in BALF. (E) Staining of lung sections shows emigrated neutrophils in the lungs (20× dry lens). (F) The number of cells in the lung capillaries was quantified as the number of cells in total capillaries (but not alveolar air spaces)/10 alveolar air space area using ImageJ. (G) Emigrated neutrophils in alveolar spaces were quantified as volume fraction of the alveolar air space using standard point-counting morphometric techniques. (H) Cytokine/chemokine levels in BALF were measured using specific ELISA kits. (I and J) G-CSF blockage–induced elevation of cytokine levels in the lungs is mediated by neutrophils. (I) Schematic representation of the experimental procedures. Neutrophils were depleted by a Gr-1 antibody in both PB (∼85% depletion) and the BM (∼65% depletion; Kwak et al., 2015). (J) Cytokine/chemokine levels in BALF. Data shown are means ± SD of three experiments (n = 5 mice). *, P < 0.01 versus control (mice treated with IgG).