Fig. 3
From: RNA inhibitors of nuclear proteins responsible for multiple organ dysfunction syndrome
In vitro efficacy of RNA aptamers. a Human platelet aggregation measurements using platelets derived from three independent healthy donors. Collagen (Col), histone H4 (H4), histone aptamers (KU7 and KU9), calf thymus histones (CTH), heparin (Hep); *p < 0.01 vs. H4, one-way ANOVA corrected for multiple comparisons (Dunnett testing), n = 3–5. b TLR activation: IL-6 ELISA detection of IL-6 protein levels (as a measurement of TLR activation) in supernatants of EA.Hy926 cells treated with media alone (Veh), calf thymus histones (CTH), histone aptamers (KU7 and KU9) alone or in combination; *p < 0.01 vs. CTH, one-way ANOVA corrected for multiple comparisons (Dunnett testing), n = 3. c Cytotoxicity measurements: aptamer inhibition of histone-mediated cytotoxicity of endothelial cells determined by MTS assay. (Left panel) EA.hy926 cells treated with 1.2 µM of aptamers (KU7 or KU9) and varying amounts (0 to 1000 µg mL−1) of calf thymus histones (CTH). (Right panel) EA.hy926 cells treated with 180 µg mL−1 of CTH and increasing amounts (0 to 16 µM) of aptamers (KU7 or KU9); **p < 0.01 vs. vehicle (100 µg mL−1), n = 4. d Dynamic changes of intracellular calcium levels in Fura 2-AM-loaded EA.hy926 cells using fluorescence microscopy. (Left panel) Representative intracellular calcium elevation traces (F340/F380) of EA.hy926 treated with CTH alone, aptamer KU7 alone (25 µg mL−1, top panels), aptamer KU9 alone (25 µg mL−1, bottom panels) or in the presence of varying aptamer amounts (Molar ratio of CTH to aptamer indicated). (Right panel) Summary of data from multiple EA.hy926 cells (n = 19–69 cells per bar); **p < 0.01 vs. CTH, one-way ANOVA corrected for multiple comparisons (Tukey's testing), n = 3. Data represent mean ± SEM
