TF cytoplasmic domain deletion enhances ERK1/2 phosphorylation in macrophages. (A) Time course shows enhanced ERK1/2, but similar p38, phosphorylation after stimulation with LPS (1 μg/mL) in TFΔCT compared to wild-type BMMs. Control experiments with confirmed TLR4-selective LPS gave similar results. (B) Nuclear translocation of phosphorylated ERK1/2 was analyzed in BMMs stimulated with LPS (1 μg/mL) for 20 minutes by triple staining of permeabilized cells with anti-phospho ERK1/2 (red), antimouse TF (green), and nuclear staining with ToPro3 (blue). (C) TF staining (green) in nonpermeabilized BMMs with and without LPS stimulation for 10 minutes.