ESAT-6 induced phosphorylation of ERK1/2 in cytoplasm but not in nucleus. 10 × 106 RAW264.7 cells were stimulated with 5 μg/ml of recombinant ESAT-6 for 0, 15, 30, 60 and 120 minutes; cytoplasmic and nuclear extracts were run on gel and probed with anti-phospho-ERK1/2 antibody. (A) phosphorylation of ERK1/2 in cytoplasm. (C) phosphorylation of ERK1/2 in nucleus. (B) and (D) Total ERK1/2 in the cytoplasmic and nuclear extracts respectively at different time points to confirm equal loading of samples in all the lanes. (E) and (G) represents phosphorylated p38 in cytoplasm and the nucleus respectively. (F) and (H) shows the total p38 protein in cytoplasm and nucleus respectively. Data is a representative from three experiments.