Open Access Research article

TNF-α Mediates Eosinophil Cationic Protein-induced Apoptosis in BEAS-2B Cells

Kun-Che Chang1, Chih-Wei Lo1, Tan-chi Fan2, Margaret Dah-Tsyr Chang2, Chih-Wen Shu1, Chuan-Hsin Chang1, Cheng-Ta Chung1, Shun-lung Fang2, Chih-Chung Chao1, Jaw-Ji Tsai3 and Yiu-Kay Lai14*

Author Affiliations

1 Institute of Biotechnology & Department of Life Science, National Tsing Hua University, Hsinchu, 30013 Taiwan

2 Institute of Molecular and Cellular Biology & Department of Life Science, National Tsing Hua University, Hsinchu, 30013 Taiwan

3 Department of Allergy and Clinical Immunology, Taichung Veterans' General Hospital, Taichung, 40705 Taiwan

4 Department of Bioresources, Da-Yeh University, Changhua, 51591 Taiwan

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BMC Cell Biology 2010, 11:6  doi:10.1186/1471-2121-11-6

Published: 20 January 2010

Additional files

Additional file 1:

Supplementary Figure 1. rECP induces DNA fragmentation in BEAS-2B cells. BEAS-2B cells (5 × 105) were incubated in a 10 cm dish in the absence or presence of 20 μM of rECP for 48 h. DNA damage indicating apoptosis was determined by the DNA fragmentation.

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Additional file 2:

Supplementary Figure 2. Effect of cytochrome c release on rECP treatment. BEAS-2B cells (5 × 104) were incubated in a 6 well plate in the absence or presence of 10, 20 μM of rECP for 24 h. The cytosolic cytochorme c was detected by western blotting.

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Additional file 3:

Supplementary Figure 3. Effects of TNF-α liberation on various RNases. BEAS-2B cells were treated with 20 μM RNase A, rECP and mECP. TNF-α was measured in cell lysates by treatment for 48 h. All the TNF-α measurements were determined by ELISA assay. All data represent the arithmetic mean ± SEM. *P < 0.05

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Additional file 4:

Supplementary Figure 4. RNase activities of recombinant eosinophil RNases degrading yeast tRNAs. The RNase activities of rECP and EDN were measured employing a standard assay with yeast tRNA as the substrate, and RNaseA as a positive control. The values indicate in nmol tRNA digested per pmol enzyme per second.

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Additional file 5:

Supplementary Figure 5. Effect of different ribonucleases on cytotoxicity of BEAS-2B cells. Equal mounts of cells were cultured in 12-well plates in the presence of 20 μM of rECP, mutant rECP, rEDN and RNase A for 48 h. The cleavage of PARP was detected by western blotting. *P < 0.05

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