TLR and HIF in Human DCs Figure 1. Time dependent stabilization of HIF-1a protein by TLR agonists, CoCl2 and Hypoxia. Monocytes isolated from buffy coat were cultured in presence of GM-CSF and IL-4 for 6 days. Cell lysates from MoDC that had been incubated with LPS (1 mg/ml, A), HA (20 mg/ml, B), LTA (5 mg/ml, C), CoCl2 (200 mM, D) or cultured under hypoxic conditions (1.5% oxygen, E) for the indicated periods were prepared. Equal amounts of protein were loaded and separated by SDS page, transferred onto membranes and probed for HIF-1a and actin. Relative Intensity Values are indicated on the right side of each blot. Protein Densiometry was quantified using Adobe Photoshop CS3. Values are representative for 3–4 experiments with cells from different donors. doi:10.1371/journal.pone.0010983.g001 PLoS ONE | www.plosone.org 3 June 2010 | Volume 5 | Issue 6 | e10983