Skip to main content
. 2013 Oct 11;1(5):e00123. doi: 10.1002/phy2.123

Figure 5.

Figure 5

Apoptosis of cIAP2-depleted airway epithelial cells (AECs) by TNFα and IFNγ correlates with cIAP1 and Bcl2 downregulation. (A) AECs from nonasthmatic donors were transfected with cIAP2-specific siRNA, and cultured for 72 h in the presence of TNFα and IFNγ (50 ng/mL). A large 19 kDa caspase-3 product is produced, with a relatively small 17 kDa product. Transfection (Txn) reagent and scrambled siRNA (sham) oligonucleotides were included to control for unintended cIAP2 silencing, and apoptosis as a result of transfection conditions. Doxorubicin (Dox) was used as a positive control for caspase-mediated apoptosis. These results are representative of experiments performed using three AEC donors. (B) Primary AECs treated with TNFα and IFNγ for 72 h exhibit downregulation of cIAP1 and Bcl2 protein abundance, while XIAP and XAF1 levels remain stable. (C) Densitometry analysis of Western blots (n = 3) shows cIAP1 and Bcl2 downregulation becomes significant when AECs are treated with 50 ng/mL of TNFα and IFNγ. Results are representative of three experiments using different AEC donors. Protein expression was baselined to the abundance in the untreated sample, and normalized to the expression of β-actin. **P < 0.01; ***P < 0.001. Error bars represent 95% confidence intervals.