LPS induced phosphorylation of ERK1/2 in both cytoplasm and nucleus. RAW264.7 cells were stimulated with 0.1 μg/ml of bacterial LPS for 0, 15, 30, 60 and 120 minutes and probed for phospho-ERK1/2 as before. (A) Phosphorylation of ERK1/2 in cytoplasm upon stimulation with LPS(0.1 μg/ml). (C) phosphorylation of ERK1/2 in nucleus. The data is a representative of three independent experiments. (B) and (D) Total ERK1/2 in cytoplasm and nucleus normalized for protein content. Costimulation of RAW264.7 cells with LPS (0.1 μg/ml) and 5 μg/ml of ESAT-6 for 0, 15, 30, 60 and 120 minutes. (E) ERK1/2 phosphorylation in cytoplasm upon stimulation with 5 μg/ml of ESAT-6 and 0.1 μg/ml of LPS. (G) Phosphorylation of ERK1/2 in the nucleus with both LPS and ESAT-6. (F) and (H) Total ERK1/2 protein in cytoplasm and nucleus respectively.