Zhu et al. Journal of Experimental & Clinical Cancer Research 2012, 31:31 http://www.jeccr.com/content/31/1/31 Page 5 of 7 Figure 2 Migration of human breast cancer cells in the presence of BLyS. 0.1 ng/ml, 1 ng/ml and 10 ng/ml BLyS were added in the lower chamber. 2% FBS and 1% FBS added in the lower chamber were used as positive chemoattractant and negative chemoattractant respectively. (A) MDA-MB-435. (B) MDA-MB-231. (C) MDA-MB-468. Average numbers of cells migrating in five randomly selected fields were counted after 8 h (original magnification 200 ×). Data are means of triplicate samples with ± SD; *, P < 0.05, **, P < 0.01, ***, P < 0.001, vs 1% FBS under normoxia. #, P < 0.05, ##, P < 0.01, ###, P < 0.001, vs 1% FBS under hypoxia. Figure 3 Role of p65 activation in BLyS up-regulation. (A) HIF-1a and p65 protein levels in MDA-MB-435 in hypoxic conditions for different time points by Western Blotting. (B) CAPE(50 μM)and PX 12 (10 μM) were used to determine the roles of p65 and HIF-1a in the regulation of BLyS expression by Western Blotting. The cells were treated with or without inhibitor in normoxic or hypoxic conditions for 6 h. (C) Effects of CAPE(50 μM)and PX 12 (10 μM) on BLyS promoter activity. Data were average luciferase activities of three independent transfections with ± SD. *, P < 0.05, vs pGL3-Basic/BP. (D) Localization of p65 protein and expression level of BLyS by immunofluorescence. MDA-MB-435 cells were challenged with CAPE (50 μM) for 6 h (original magnification 200 ×).