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. 2012 Aug 22;7(8):e43459. doi: 10.1371/journal.pone.0043459

Figure 6. Proteasome inhibition prolongs the pSmad3 signal with a dependence on TβRI kinase activity.

Figure 6

A, α-pSmad3C, α-tSmad3 and α-clathrin immunoblots of ES-2 cells, treated with 2ME2 or vehicle, proteasome inhibitors (MG132 and ALLN), or their combination, and stimulated with TGF-β1 for the indicated times. B, Bar graph depicts average ± SD of the proteasome inhibitor protection factor (PIP) at each time point after ligand addition. The factor is defined as PIP = 1−(pSmad3C/tSmad3/clathrin)uninhibited/(pSmad3C/tSmad3/clathrin)inhibited. n = 3; p<0.015, 1-tailed t-test. C, α-pSmad3C, α-tSmad3 and α-clathrin immunoblots of ES-2 cells, treated with proteasome inhibitors or vehicle and stimulated with TGF-β1. In indicated samples, SB431542 was added at 1 h after TGF-β1 stimulation.