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Author: Bond, D
Author: Foley, E
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Methods Article

Quantitative Evaluation of Signaling Events in Drosophila S2 Cells

David Bond1, David A. Primrose1 and Edan Foley1*

1 Department of Medical Microbiology and Immunology, University of Alberta. Edmonton AB. Canada.

* To whom correspondence should be addressed: Edan Foley, Department of Medical Microbiology and Immunology, University of Alberta. Edmonton AB. Canada. Phone: (780) 492-2303. Fax: (780) 492-9828. Email: efoley@ualberta.ca

Biol. Proced. Online 2008;10:20-28. doi:10.1251/bpo139
Submitted: September 24, 2007; Accepted: December 09, 2007; Published: February 04, 2008.

Indexing terms: RNA interference; immunity; drosophila.

Abbreviations: IKK, I-kappa kinase; Imd, immune deficiency; JNK, c-Jun N-terminal kinase; PGN, gram-negative bacterial peptidoglycan; PGRP-LC, peptidoglycan recognition protein-LC; TNF, tumor necrosis factor.


Figure 1 Enlarged

Fig. 1:

Time course of Bsk phosphorylation in response to LPS. (A,B) Western blot analysis of total and phospho-Bsk protein (p-Bsk) in S2 cells treated with LPS for the indicated periods. Lysates were probed with anti-p-JNK (A) and anti-JNK (B) antibodies on a single blot. The primary antibodies were detected with Alexa-fluor 680 and Alexa-fluor 750-labeled secondary antibodies, respectively. Panels A and B are false-colored and merged in C, with total Bsk in red and p-Bsk in green. Molecular mass markers are shown in lane 9. (D) p-Bsk protein levels in (A) were quantified and normalized relative to total JNK protein levels in (B) for each time point. The p-JNK:total JNK ratio at 0min LPS exposure was assigned a value of one and all other ratios are shown relative to this value. Treatment of S2 cells with LPS led to a transitory phosphorylation of Bsk with maximal Bsk phosphorylation at 5 mins.

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