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. Author manuscript; available in PMC: 2010 Aug 1.
Published in final edited form as: Am J Reprod Immunol. 2009 Aug;62(2):96–111. doi: 10.1111/j.1600-0897.2009.00717.x

Figure 2. Effects of anti-β2GPI Abs on trophoblast cell viability and apoptosis.

Figure 2

(A) First trimester trophoblast cells (HTR8) were incubated with ID2, IIC5 or the mouse IgG1 isotype control (IgG) at 0, 5, 10, 20 and 40µg/ml for 48 hours. Cell viability was then determined using the CellTiter 96 assay. Line graph shows the percentage cell viability relative to the untreated control (0 µg/ml). Treatment with ID2 or IIC5 at the high dose of 40µg/ml significantly reduced trophoblast cell viability when compared to the untreated control (*p<0.05, **p<0.001). This figure is representative of at least three independent experiments performed in triplicate. Significance was determined using ANOVA.

(B) First trimester trophoblast cells (HTR8) were incubated with no treatment (NT); ID2 (40µg/ml); IIC5 (40µg/ml); mouse IgG (40µg/ml), as a negative isotype control (IgG); or CPT, (4µM) as a positive control for cell death, for 48 hours. Cell viability was then determined using the CellTiter 96 assay. Barchart shows the percentage cell viability relative to the NT control *p <0.05, **p <0.001. Significance was determined using ANOVA. Data are pooled from five individual experiments.

(C) First trimester trophoblast cells (HTR8) were incubated with NT, ID2 (40µg/ml); IIC5 (40µg/ml) or mIgG (IgG; 40µg/ml) in the presence and absence of heparin (100ng/ml) for 48 hours. Cell viability was then determined using the CellTiter 96 assay. Barchart shows the percentage cell viability relative to the NT control unless otherwise indicated and significance was determined using ANOVA. The presence of heparin significantly reduced the amount of cell death induced by ID2 and IIC5 (*p <0.01, **p <0.001), as determined using a paired t-test. Data are pooled from three individual experiments.

(D) Trophoblast cells (HTR8) were incubated with either no treatment (NT), ID2 (40µg/ml) or IIC5 (40µg/ml). After 48 hours, caspase-3; caspase-8; and caspase-9 activation was determined. Barchart shows caspase activity in relative light units (RLU) relative to the NT control (*p<0.001). Significance was determined using ANOVA. Data are pooled from three individual experiments.