Figure 7.

Effect of HIF-1α siRNA on the xenon mediated cytoprotection. HK-2 cells were treated with either scrambled siRNA or HIF-1α siRNA (20 nM) for 6 h, followed by xenon gas (70% Xe and 5% CO₂ balanced with O₂) for 2 h and then recovered in normal cell incubator for 24 h. (A) Immunoluorescent dual-labeling of HIF-1α (Red) and VEGF(Green) in HK-2 cells. Scale bar: 50 μm. (B) Knock-down of HIF-1α was confirmed by Western blot. (C) Mean fluorescence intensity of HIF-1α and VEGF after HIF-1α siRNA treatment. The cells underwent 24 h hypothermia-hypoxia challenge and recovered (reoxygenation) for 24 h. (D) Effect of xenon on HMGB-1 (Red) cytoplasmic translocation, was abolished by HIF-1α siRNA treatment. Scale bar: 50 μm. Percent of HK-2 cells with cytoskeleton disorganization (E), nuclear fragmentation (F) and HMGB-1 cytoplasmic translocation (G) per 20× images. (H) Cell viability was assessed by an MTT assay. Data is expressed as mean ± SD (n = 4); *p < 0.05 and ***p < 0.001.