BDNF induces ERK1/2 and AKT phosphorylation in BMSCs, but has no effect on NF-κB activation.

<p>(A, B, C) BMSCs were cultured in the presence of 25 ng/ml BDNF for 5, 15, 30, or 60 min. The expression of p-ERK1/2 (A) and p-Akt (B) began to increase at 5 min. However, no obvious change in the I-κB (C) level was detected by 60 min after BDNF stimulation. The results are representative of three independent experiments. Statistical analysis was conducted by ANOVA, * <i>P</i><0.01 versus controls. (D) BDNF does not induce translocation of NF-κB p65 to the nucleus in BMSCs. BMSCs were treated with BDNF for 5, 15, 30 or 60 minutes, then fixed and probed with antibodies specific for NF-κB p65. Magnification ×200. Representative images from 15-min exposures to stimuli are exhibited. (E) MEK1/2 inhibitor U0126 and PI3K inhibitor LY204002 blunted the increase of RANKL protein secretion induced by BDNF. The results represent the means ± SEMs for all assays (performed in triplicate) (BDNF+U0126 vs. BDNF, * <i>P</i><0.05; BDNF+LY204002 vs. BDNF, <i>P</i> = 0.069; BDNF+U0126+LY204002 vs. BDNF, ** <i>P</i><0.05).