Fig. 2:

In-Cell Quantification of Bsk Phosphorylation. (A,B) In-Cell quantitative analysis of S2 cells or S2 cells incubated with TAK1 or Kenny dsRNA and treated with LPS for the indicated periods. Cells were incubated with anti-p-JNK antibodies. The primary antibody was detected with an Alexa-fluor 750-labeled secondary antibody (A). Cellls were counterstained with Alexa-fluor 680-labeled phalloidin in (B). Panels A and B are false-colored and merged in (C) with p-Bsk in green and f-actin in red. (D) p-Bsk protein levels from (A) were quantified and normalized to f-actin levels in (B) for each time point. The p-JNK:total JNK ratio at 0min LPS exposure was assigned a value of one for each row and all other ratios are shown relative to this value. Error bars represent the standard error of two independent experiments. Treatment of S2 cells with LPS led to a transitory phosphorylation of Bsk, which was eliminated by dsRNA-mediated depletion of dTAK1 and enhanced and prolonged by dsRNA-mediated depletion of Kenny.