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. 2014 Nov 6;6(1):43–55. doi: 10.18632/oncotarget.2723

Figure 2. Syringaresinol protects cardiomyocytes against H/R-induced apoptosis.

Figure 2

H9c2 cells were exposed to hypoxia stress for 16 h and treated with 25 μM syringaresinol during reoxygenation. After 9 h of reoxygenation, cell death was determined by TUNEL staining and flow-cytometric analysis (A) Representative images of TUNEL-positive cells (green) and Hoechst 33342 counterstaining (blue). Scale bar: 50 μm. (B) The relative proportion of TUNEL-positive cells. (C) Flow cytometry analysis of the annexin V-FITC/PI staining. (D) Quantification histograms indicate the percentages of early (annexin V-FITC positive and PI negative) and late (annexin V-FITC positive and PI positive) apoptotic cells. (E) Expression levels of apoptotic-related proteins, BCL-2 and BAX were examined by Western blot. Actin levels are shown as loading controls. (F) BCL-2/BAX ratios were assessed. (G) Caspase-3 activity was detected using a commercial kit as described in Materials and Methods. Results are presented as fold-changes over the control group. All results are representative or means ± SD of six independent experiments. Ctrl: control; H/R: hypoxia/reoxygenation; SYR: syringaresinol. #P < 0.001 versus control group, *P < 0.001 versus H/R treatment group.