TNFα promotes increased MAP4K4 protein expression through the JNK pathway.

<p><b>A</b>) IB analysis of cells treated for 24 h with increasing concentrations of TNFα using antibodies against MAP4K4 or tubulin; Below: quantification of integrated densities of bands (means −/+ SDs, normalized to tubulin). <b>B</b>) Time course IB analysis of lysates of cells treated with 25 ng/ml TNFα using antibodies against proteins indicated. Right: IB analysis of effect of JNK inhibitor SP600125 (10 ng/ml, added 2 h before TNFα treatment) on TNFα-induced JNK pathway activation. Bar diagram: Means and SDs of pJNK and pATF2 after TNFα stimulation (normalized to total JNK and total ATF2 proteins, respectively). <b>C</b>) Time course IB analysis of lysates of cells treated with 25 ng/ml TNFα using antibodies against proteins indicated. Cells were stimulated 24 h after transfection with siControl or siMAP4K4_1 or siMAP4K4_2. Bar diagram below shows quantification of pATF2 bands (normalized to total ATF2 protein). <b>D</b>) IB analysis of lysates from cells treated for 24 h−/+25 ng/ml TNFα and −/+10 ng/ml SP600125, using anti-MAP4K4 or anti-tubulin antibodies. Bar diagram below shows quantification of MAP4K4 bands (means −/+ SDs, normalized to tubulin).