Fig 11. Transfection of MAP3K19 renders HEK cells susceptible to Compound A-mediated inhibition of P-Smad2 nuclear translocation.

HEK cells, which express very low levels of MAP3K19 as determined by RT-qPCR analysis, were transfected with either empty vector (pcDNA) or MAP3K19. Following an overnight culture, the cells were pretreated with Compound A (1 μM) for 30 minutes and treated with TGF-β1 (1 ng/ml) for one hour. The cells were then harvested, and the nuclear lysates were assayed by Western analysis for nuclear phospho-Smad2 localization. The blot was then re-hybridized with HDAC-1 to show equal protein loading. Densitometric analysis of P-Smad2/HDAC-1 levels from the Western blot is shown. This experiment is representative of two independent experiments.