Ser276 Phosphorylation of NF-kB p65 by MSK1 Controls SCF Expression in Inflammation (original) (raw)
Figure 2
Effect of MAP kinase and MSK1 inhibition on IL-1β-induced NF-κB activation.
A. Human lung fibroblasts in culture were pre-incubated for 1 h with a combination of SB202190 (SB; 3.5 µM) and PD98059 (PD; 20 µM) and treated with IL-1β for the indicated time. Western blot analysis used anti-phospho-Ser276-p65, anti-phospho-Ser536-p65, anti-IκB and anti-p65 antibodies. B. Cells were transfected with 1 µg of pHis-p65 plasmid (WT p65 subunit with a N-terminal His-Tag) and pre-incubated with a combination of SB and PD, or H89 (10 µM) for 1 h before IL-1ß treatment. C. Cells were co-transfected with His-p65 plasmid and WT or “kinase-dead” (KD) MSK1 plasmid (1 µM), and treated with IL-1β (20 U/ml). Results are representative of three independent experiments performed in fibroblasts from three different donors.