Inflammatory Monocytes Orchestrate Innate Antifungal Immunity in the Lung
Figure 5
Inflammatory responses of CCR2+Mo and Mo-DC during respiratory fungal infection.
Lung CCR2+Mo (GFP+CD45+CD11b+CD11c−Nk1.1−) and Mo-DC (GFP+CD45+CD11b+CD11c+NK1.1−) were FACS sorted 48 h p.i. from CCR2 reporter mice (purity >97% for all sorts) for transcriptome analysis by RNA-seq (A) or for quantitative RT-PCR (B). Control CCR2+Mo were also isolated from the lung of uninfected CCR2 reporter mice (naïve sample) to >97% purity. (A) Gene expression data shown in A is for one experiment and representative of 3 independent biological replicates and three idependent sequencing reactions using SOLiD sequencing platform. Differences in gene expression are shown as fragments per kilobase (FPKM) as calculated using Cufflinks and R software. (B) The graphs show expression of specific transcripts in the indicated cell populations by qRT-PCR using Taq-Man probes normalized to GAPDH. Data shown is mean ±SEM pooled from two separte experiments. (C) The graph shows pulmonary Nos2 induction in DT-treated CCR2 depleter and control mice at the indicated time points p.i. Data shown is mean ±SEM pooled from two separte experiments with 3 mice per group per time point. (D–E) The scatterplots show mean ± SEM lung (D) IL-12p70 and (E) TNF levels at 48 h p.i. in CCR2 depleter (grey circles) and control B6 mice (black circles) as in Figure 3A.