Figure 1. Effect of Tβ4 on cell viability in H₂O₂ treated fibroblasts.

(A) The MTT assay was performed with increasing H₂O₂ concentration (1 to 250 µM) in presence (dotted lines) and absence (solid lines) of Tβ4 (1 µg/mL). Data represent means±SEM of 3 individual experiments. (B) Representative confocal laser scanning microscopy images of cardiac fibroblasts stained with DCF-DA showing the effect of Tβ4 on intracellular ROS upon treatment with H₂O₂. (C). Effect of Tβ4 on generation of ROS in fibroblasts treated with H₂O₂ by fluorimetry. The graph represents the percentage of fluorescence positive fibroblasts upon staining with DCF-DA. Data represent the mean±SE of at least three separate experiments. * means p<0.05 compared to the controls and # represents p<0.05 compared to the respective H₂O₂ treated group (D) Representative confocal laser scanning microscopy images of cells stained with DHE Red showing the effect of Tβ4 on generation of superoxide radicals upon treatment with H₂O₂ in fibroblast. (E). Representative confocal laser scanning microscopy images of cells stained with DAF-2DA showing the effect of Tβ4 on generation of nitric oxide upon treatment with H₂O₂ in fibroblast. (F). Representative confocal laser scanning microscopy images of cells stained with Mitotracker Red showing the effect of Tβ4 on loss of mitochondrial membrane potential upon treatment with H₂O₂ in fibroblast.