Hypoxia-inducible factor-1α (HIF-1α) in patients with sepsis and healthy volunteers. A, N-fold change in HIF-1α mRNA expression in patients with sepsis compared to healthy volunteers. Specific mRNA expression was analyzed using real-time polymerase chain reaction. Data were analyzed using the Δct method and are depicted as n-fold induction (2^ΔΔct, mean ± SEM). In patients with sepsis, HIF-1α mRNA expression was decreased by 67% compared to that in healthy controls. * P = 0.0001. B, Intracellular HIF-1α protein was stained by immunofluorescence in blood smears from patients with sepsis and healthy volunteers. Blood smears were performed immediately after blood sampling (unstimulated cells) and following 2h of incubation with dimethyloxalylglycine (10 µm). First, representative examples of blood smears from healthy volunteers and patients with sepsis at 40-fold magnification and of negative controls (secondary antibody only) are depicted. Representative examples of erythrocytes, and HIF-1α-negative, intermediate, and positive leukocytes are shown below. HIF-1α protein was lower in unstimulated cells of patients with sepsis and even following HIF-1 protein stabilization using dimethyloxalylglycine. In negative controls (secondary antibody only), there was no unspecific antibody binding. DMOG = dimethyloxalylglycine; mRNA = messenger ribonucleic acid.