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. 2013 Jul 9;8(7):e67990. doi: 10.1371/journal.pone.0067990

Figure 5. PI3K/Akt pathway is involved in BDNF-induced migration and β5 integrin up-regulation in human chondrosarcoma cells.

Figure 5

(A) Cells were incubated with BDNF (50 ng/ml) for the indicated time intervals, and p-p85 and p-Akt was examined by western blotting (n = 5). (B) Cells were pretreated for 30 min with Ly294002 (10 µM), wortmannin (1 µM), and Akt inhibitor (1 µM), or transfected with dominant negative (DN) mutants of p85 and Akt for 24 h, and then allowed to migrate for 24 h toward BDNF (50 ng/ml)-containing medium. In vitro migration was examined using a Transwell assay (n = 4). (C&D) Cells were pretreated for 30 min with Ly294002 (10 µM), wortmannin (1 µM), and Akt inhibitor (1 µM), or transfected with dominant negative (DN) mutants of p85 and Akt for 24 h, followed by stimulation with BDNF. The mRNA and cell-surface β5 integrin expression was examined using qPCR and flow cytometry (n = 5). Cells were pretreated with K252a and TrkB mAb (E) or K252a and Ly294002 (F) for 30 min. This treatment was followed by stimulation with BDNF (50 ng/ml) for 15 min, and p85 (E) and Akt (F) phosphorylation were examined (n = 4). Results are expressed as the mean ± SEM. *, p<0.05 compared with the control. #, p<0.05 compared with the BDNF-treated group.