Figure 4.

Deferoxamine (DFO) modulates the expression of Wnt5a via the PI3K and NFAT pathways. (A) Day 10 differentiated bone marrow stromal cells (BMSCs) were treated with 50 μM DFO for 30 min. Western blot analysis was performed for various signaling pathways. Protein amounts were normalized to GAPDH. Three independent experiments were performed. Two are shown. (B–C) BMSCs were treated with the following inhibitors prior to the addition of 50 μM DFO for 48 hours: BAY-11-7082 (I-κB phosphorylation inhibitor, 1 μM), UO126 (MEK1 inhibitor, 10 μM), LY-294002 (PI3K inhibitor, 10 μM), and VIVIT (NFAT inhibitor, 1 μM). Afterwards, Wnt5a expression was determined using either real-time polymerase chain reaction (PCR) (B) or Western Blot analysis (C). All three Western blots were quantified using ImageJ. N=3–4. *P<0.05 vs. control (CO).