Figure 4.

Clusterin prevents cell apoptosis and death induced by I/R in vitro. H9C2 cells (80,000 cells/ well) were plated in six well plates and allowed to culture at 37°C 5% CO₂ overnight. Cells were infected with human clusterin expression adenovirus or control null virus for 24 hours. Cells were then subjected to a hypoxia chamber with 0% O₂ and 15% CO₂ at 10°C for 16 h, followed by 24 hours reperfusion at 5% CO₂, 28% O₂ at 37°C. A. Cell apoptosis was detected by TUNEL staining; B. Cell apoptosis was detected by double staining with FITC labeled Annexin-V and PI and flow cytometry; C. Protein expression of clusterin, bax, and bcl-XL was detected by Western blot assay; D. Gene expression of clusterin, bax, and bcl-XL was detected by RT-PCR; E. Gene expression of clusterin, bax, and bcl-XL was detected by qRT-PCR. Data are the summary of three independent experiments. Vs. I/R, *P<0.01