MSK1 interacted with RelA/p65 and p300.

<p>(A) H292 cells were transiently transfected with wild-type MSK1 or MSK1 N-terminal kinase-dead mutant MSK1 flag construct and treated with or without CSE (1.0%) for 1 h. Flag-tagged precipitated complexes were analyzed through immunoblotting using anti-flag and anti-RelA/p65 antibodies. Whole cell lysate from pCMV-Flag-MSK1 wild-type transfected cells were used as lysate control. (B) H292 cells were treated for 1 h with or without CSE (2.0%) and TNFα (10 ng/ml) was used as positive control. Nuclear extract (1000 µg) was used for immunoprecipitation of MSK1 bound complexes and analyzed through immunoblotting using anti-MSK1, anti-RelA/p65 and anti-p300 antibodies. (C) H292 cells were treated for 1 h with or without CSE (2.0%), and TNFα (10 ng/ml) was used as positive control. Nuclear extract (1000 µg) was used for immunoprecipitation of RelA/p65 bound complexes and analyzed through immunoblotting using anti-RelA/p65 and anti-MSK1 antibodies. (D) CS-mediated interaction of MSK1-p300 in acute CS exposed mouse lung. Adult C57BL/6J mice were exposed to CS at 300 mg/m³ total particulate matter (TPM) for 3 days and were sacrificed 24 h after the final exposure. Whole tissue extract (500 µg) was used for immunoprecipitation of MSK1 bound complexes, and analyzed by immunoblotting using anti-MSK1 and anti-p300 antibodies. (E) CS-mediated interaction of RelA/p65 with MSK1 and p300 in acute CS exposed mouse lung. Adult C57BL/6J mice were exposed to CS at 300 mg/m³ total particulate matter (TPM) for 3 days and were sacrificed 24 h after the final exposure. Whole tissue extract (500 µg) was used for immunoprecipitation of RelA/p65 bound complexes, and analyzed by immunoblotting using anti-RelA/p65, anti-MSK1 and anti-p300 antibodies. Results are representative of at least three separate experiments.