Fig 2: Impact of reduced OPG and RANK expression in MDA-MB-231 cells on cell-matrix adhesion in vitro. Reduced OPG expression did not alter MDA-MB-231 cell-matrix adhesion compared with control cells (A). Reduced RANK expression resulted in a significant decrease in MDA-MB-231 cell-matrix adhesion compared with control cells (B). When MDA-MB-231pEF6 control cells were treated with 40 ng/ml HGF or 50 µg/ml BME or a combination of 40 ng/ml HGF and 50 µg/ml BME did not significantly alter MDA-MB-231 cell-matrix adhesion (C). MDA-MB-231OPGKD cells treated with 40 ng/ml HGF or 50 µg/ml BME also did not alter cell-matrix adhesion, however, a combined of HGF and BME treatment significantly decreased cell matrix adhesion (D). MDA-MB-231RANKKD cells treated with 40 ng/ml HGF, 50 µg/ml BME or combined HGF and BME did not affect cell-matrix adhesion compared to the untreated cells (E). Data represent the mean of 4 independent repeats, error bars represent SEM. *p=0.05.

Fig 3: Impact of reduced OPG and RANK expression on MDA-MB-231 cell invasion in vitro. MDA-MB-231OPGKD cells showed significantly increased cell invasion compared with MDA-MB-231pEF6 control cells (A). MDA-MB-231RANKKD cells showed reduced cell invasion compared with MDA-MB-231pEF6 control cells (B). Treatment of MDA-MB-231pEF6 control cells with 40 ng/ml HGF, 50 µg/ml BME or a combination of 40 ng/ml HGF and 50 µg/ml BME do not impact cell invasion (C). Treatment of MDA-MB-231OPGKD cells with 40 ng/ml HGF or a combination of 40 ng/ml HGF and 50 µg/ml BME resulted in a significant decrease in cell invasion (D). MDA-MB-231RANKKD cells treated with 40 ng/ml HGF, 50 µg/ml BME or combined 40 ng/ml HGF and 50 µg/ml BME showed non-significant increases in cell invasion (E). Data represent the mean of 3 independent repeats, error bars represent SEM. *p==0.05, **p=0.01.

Fig 4: Impact of OPG and RANK knockdown on MDA-MB-231 cell proliferation in vitro. Suppression of OPG expression had no effect on MDA-MB-231 cell proliferation after 5-day incubation compared to MDA-MB-231pEF6 control cells (A). Reduced RANK expression in MDA-MB-231 cells resulted in a significant decrease in cell proliferation after 5-day incubation compared with control cells (B). Treatment of the MDA-MB-231pEF6 control cell line with 40 ng/ml HGF, 50 µg/ml BME or a combination of 40 ng/ml HGF and 50 µg/ml BME resulted in a significant increase in cell proliferation after 5-day incubation compared to the untreated cells (C). After 5-day treatment of MDA-MB-231OPGKD cells or MDA-MB-231RANKKD cells with 40 ng/ml HGF, 50 µg/ml BME or a combination of 40 ng/ml HGF and 50 µg/ml BME no changes were seen compared to the respective untreated cells (D and E). Data represent the mean of 4 independent repeats, error bars represent SEM. *p=0.05.

Fig 5: Plasma levels of Weibel-Palade body constituents in Indonesian children with dengue.(A) VWF antigen, (B) VWF propeptide and (C) osteoprotegerin levels (all determined by ELISA) in Indonesian children with dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS) and in healthy controls. Horizontal lines represent median values. The upper limit of detection of the OPG assay was 1200 pg/ml; 49% and 70% of children in the DHF and DSS group, respectively, had an OPG plasma level above this cut-off value at enrollment. No plasma available for analysis in two patients from the DHF group and one from the DSS group at day 1 and in fifteen and six patients from the DHF and DSS group at discharge. P values were determined by Wilcoxon matched-pairs signed rank test for data in time and Mann Whitney U- test for comparison with the control group. *p value<0.05, **p value<0.01.