(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.

Copyright information:

Taken from "Quantitative Evaluation of Signaling Events in Drosophila S2 Cells"

Biological Procedures Online 2008;10():20-28.

Published online Jan 2008

PMCID:PMC2275046.

Article © by the author(s). This paper is Open Access and is published in Biological Procedures Online under license from the author(s). Copying, printing, redistribution and storage permitted. Journal © 1997-2008 Biological Procedures Online.