1. Identification of major cellular proteins synthesized in response to interleukin-1 and interleukin-6 in human hepatoma HepG2 cells
- Author
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Bonar, Emilia, Dubin, Adam, Bierczyńska-Krzysik, Anna, Noga, Marek, Silberring, Jerzy, Stalińska, Krystyna, and Koj, Aleksander
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INTERLEUKIN-1 , *INTERLEUKIN-6 , *POLYACRYLAMIDE gel electrophoresis , *GEL electrophoresis , *INTERLEUKINS - Abstract
Abstract: Interleukin-1 (IL-1) and interleukin-6 (IL-6) are principal proinflammatory cytokines inducing the acute phase response of various tissues, including liver. Cultured human hepatoma HepG2 cells were stimulated with IL-1 (10ng/ml) and IL-6 (10ng/ml). After 24h the cells were collected and disrupted by sonication in a lysis buffer containing 8M urea. The extracted cellular proteins were separated by 2D polyacrylamide gel electrophoresis. The gels were stained with Coomassie Brilliant Blue R-250 and the protein spots showing different intensities in comparison to control (unstimulated) cells were excised and subjected to analysis by LC-MS/MS. Alternatively, proteins were stained with SYPRO Ruby. These differentially expressed proteins include seven up-regulated and two down-regulated intracellular proteins of various functions. The identification of three cytokine-responsive proteins was confirmed by biosynthetic labeling with [35S]methionine after incubation of HepG2 cells, and by western blot with specific antisera. [Copyright &y& Elsevier]
- Published
- 2006
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