Fig 8. MAP3K19 promoted the nuclear accumulation of the activated R-Smads phospho-Smad2 and phospho-Smad3 following TGF-β1 stimulation.

(A) HeLa cells were transfected with nonsense siRNA (250 or 500 ng) or MAP3K19 siRNA (250 or 500 ng). Eighteen hours later, the cells were treated with TGF-β1 (1 ng/ml) for 1 hour, harvested and nuclear extracts were isolated. Nuclear lysates were examined by Western analysis for P-Smad2, P-Smad3 and Smad4 levels, and the blots were re-probed with HDAC-1 to show equal protein loading. Densitometry was performed comparing either the P-Smad2/HDAC-1 levels or the P-Smad-3/HDAC-1 levels. Smad4 levels were not examined by densitometry, as they were not affected. This experiment was repeated three independent times and a representative experiment is shown.