Figure 3.

PHLDA1 attenuates the activation of some signal molecules in MyD88-dependent TLR4 signaling pathway. (A) RAW264.7 cells were transfected with EV or PHLDA1 plasmid, and then stimulated with LPS (0.1 μg/ml) for the indicated times. Phosphorylation levels and total protein expressions of important signal molecules (ERK, JNK, p38, IKKα/β, IkBa and p65) in cell lysates were analyzed using Western blot. (B) RAW264.7 cells were transfected with Control siRNA or PHLDA1 siRNA1, and then stimulated with LPS (0.1 μg/ml) for the indicated times. Phosphorylation levels and total protein expressions of the above molecules were analyzed using Western blot. Data shown are presentative of three independent experiments. Phosphorylation levels of the above molecules were quantitated and shown in the right panel. GAPDH was used as a loading control.