Ethyl Acetate Fraction of Hedyotis diffusa Willd Induces Apoptosis via JNK/Nur77 Pathway in Hepatocellular Carcinoma Cells

Figure 5

EHDW induced apoptosis in Hep3B cells. (a) Cell apoptosis was measured using flow cytometry. Representative data were analyzed with Annexin-V/PI staining in Hep3B cells after 24 h of exposure to indicated concentrations of EHDW. (b) Flow cytometry analysis of the percentage of apoptotic cells (early apoptosis and late apoptosis) from three independent experiments. and vs. control. (c) Apoptotic morphological observation. Fluorescence microscope images showed morphological changes in Hep3B cells upon 150–250 μg/mL EHDW treatment for 24 h White arrows showing bright blue regions indicated nuclear pyknosis and fragmentation of chromatin. (d) Expression of cyto c in cytosol and PARP, cleaved-PARP, Bcl-2, BAX, cleaved-caspase 3 in total proteins were examined by Western blotting. , , and vs. control. One-way ANOVA, post hoc comparisons, Tukey’s test. Columns, means, error bars, SEM. (e) EHDW induced apoptosis in vivo. The 5-μm zebrafish tissue sections were deparaffinized and stained with Hoechst 33258 dye. Representative fluorescent photographs were taken by fluorescence microscopy.

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