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. 2011 Oct 17;121(11):4548–4566. doi: 10.1172/JCI57734

Figure 11. Inhibition of calpain blocks EGF- and PDGF-BB–induced increases in intracellular content of active TGF-β1 and p-Smad2/3 in PASMCs.

Figure 11

PASMCs were incubated with EGF (10 ng/ml) or PDGF-BB (10 ng/ml) in the presence and absence of MDL28170 (20 μM) for 2 hours, after which the intracellular content of active TGF-β1 (A) and p-Smad2/3 (D, E, H, and I) was measured using Western blot analysis; intracellular active TGF-β1 content was also assayed using ELISA (B and C). (F and G) PASMCs were transfected with an siRNA against the mRNA of calpain-1 or calpain-2 or control (luciferase) siRNA. After 96 hours, the cells were incubated with EGF (10 ng/ml) for 2 hours, and then the protein content of p-Smad2/3 and total Smad2/3 was measured using immunoblotting. (A, D, F, and H) Representative immunoblots from 4 experiments. (E, G, and I) Changes in p-Smad2/3 and total Smad2/3 content quantified by scanning densitometry. Results are expressed as mean ± SEM; n = 4 experiments. *P < 0.05, **P < 0.01 versus control (without EGF or PDGF-BB). #P < 0.05 versus vehicle group in control siRNA. MDL, MDL28170.