Fig. 2
Transcriptional and physiological profiling of HD human monocytes. The transcriptome of fresh and HD human monocytes cultured for 2, 10 or 24 h was investigated using microarray analysis. a, Pre-ranked GSEA; genes were ranked according to their differential expression between monocytes at 2, 10 or 24 h post-HD culture and fresh monocytes. Twenty Hallmark gene sets (v7.2) were significantly overrepresented (FDR < 0.05). Upregulated gene expression is signified in red and downregulation in blue. b, Enrichment plot of the Winter hypoxia gene set in monocytes post-HD culture. c, Heat map of FCGR gene expression. d, Microarray gene expression data was acquired using fresh monocytes (M0) and monocytes at 2 (M2),10 (M10) and 24 (M24) hours post-HD culture as well as monocytes cultured under hypoxic conditions (1% O2) for 24 h (Bosco et al., 2006). Heat map of activation z-scores for the top 50 genes and proteins determined to be the most activating or inhibiting. e, % O2 in LD and HD cultures of human PBMCs and isolated monocytes (n = 6 per group, thickness of lines for LD and HD represent SEMs for each time point). f, pH and Lactate in donor matched LD and HD PBMC culture supernatants (n = 5 per group). g, LD and HD monocyte cell lysates were generated and HIF-1α and HSC70 expression assessed using Western blotting. Representative Western blot staining for 2 donors is shown. h-i, Representative histograms and graphs showing expression of HIF-1α, CAIX and GLUT1 expression quantified using flow cytometry of LD and HD precultured monocytes (n = 11 per group). Each point on the graphs represents a unique healthy donor. Statistical significance between groups was assessed by using a paired two-tailed Wilcoxon test (*p < 0.05, **p < 0.001, ***p < 0.001 and ****p < 0.0001). Also see Additional file 2: Fig. S2