Fig. 5

PepN-pretreatment preprograms CD11c^high macrophages for an OXPHOS preference and an anti-inflammatory phenotype. a Principal component analysis of RNA-seq data of CD11c^high macrophages. b Genes significantly (p adjusted < 0.05) up-regulated and down-regulated are depicted in red and green, respectively. c GO biological process enrichment results. d KEGG pathway enrichment results. e Gene set enrichment analysis of the RNA sequencing data. f–h Heat map diagrams of relative expression levels of mitochondrial respiratory chain complex-related genes (f), positive regulation of wound healing-related genes (g) and cell death in response to oxidative stress-related genes (h). i Transmission electron photomicrographs depicting mitochondrial morphology of CD11c^high macrophages. Scale bars, 2 μm and 500 nm. j Mitochondrial morphology (Mito Tracker) and mt-ROS production (Mito SOX) of CD11c^high macrophages observed using confocal microscopy. Scale bars, 25 μm. k, l Mitochondrial membrane potential (k) and mt-ROS (l) of CD11c^high macrophages were detected by flow cytometry. m Oxygen consumption rate (OCR) of CD11c^high macrophages with indicated treatments (n = 3). N Enzymatic activities of mitochondrial complex I of CD11c^high macrophages (n = 3). o ATP production by CD11c^high macrophages (n = 3). p Activation of AMPK, STAT3 and NF-κB signaling pathway in CD11c^high macrophages were detected by Western blotting. q IL-1β levels in CD11c^high macrophages measured by Western blotting. r Cytokines in supernatants of CD11c^high macrophages after restimulation for 1 d ex vivo with HDM (1 μg/ml) were measured by ELISA. Statistical analysis by a two-tailed Student’s t test (n, o) or one-way ANOVA with Tukey’s multiple-comparison test (r). Data represent one of at least five samples from three independent experiments (p, q). Data are represented as mean ± SD (n, o and r). ns, not significant