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. 2014 Aug 27;9(8):e106155. doi: 10.1371/journal.pone.0106155

Figure 2. Effect of mTOR inhibitors on TGF-β activation of mTOR and Smad pathways.

Figure 2

Serum-deprived IPF fibroblasts were treated with TGF-β for 60 minutes or left untreated in (A), followed by Western blot analysis with anti-phospho Akt (Ser473 or Thr 308) and anti-total Akt antibodies, or in (B) for 6 hours in the presence or absence of indicated inhibitors MLN0128 (0.2 µM), PP242 (2 µM), or rapamycin (0.02 µM), followed by Western blot analysis with anti-phospho-S6 and anti-α-tubulin antibodies. (C) Serum-deprived IPF fibroblasts were treated with or without TGF-β for 15 minutes in the presence or absence of indicated inhibitors followed by Western blot analysis with an anti-phospho-Smad2 or Smad3 antibody. Expression of total Smad-2, 3, 4 and 7 was analyzed by Western blot. Experiment was done on three lines, which are shown in Figure 1; results were similar between the three lines and results from the IPF fibroblasts isolated from surgical lung biopsy are shown here.