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.
Taken from "Quantitative Evaluation of Signaling Events in Drosophila S2 Cells"
Biological Procedures Online 2008;10():20-28.
Published online Jan 2008
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