Skip to main content
. Author manuscript; available in PMC: 2012 Dec 1.
Published in final edited form as: Clin Cancer Res. 2011 Sep 26;17(23):7424–7439. doi: 10.1158/1078-0432.CCR-11-1248

Figure 5.

Figure 5

(A) A549 cells stably expressing the BTR WT and MUT reporter were treated with increasing concentrations of RTK inhibitor Lapatinib (0.1, 1, 2, 5, and 10µM) and Tyrphostin AG1478 (B). Bioluminescence was measured after 1hr. The change in bioluminescence activity over mock treatment (DMSO) levels was calculated and plotted as fold induction. Bioluminescent measurements were done in 8 wells. (C) A549 cells were treated with TGFβ (10ng/ml) or TGFβ and SB431542, PP2, AG1478 and Lapatinib (10 µM) for 1 h, cell lysates were prepared and immunoprecipitated with Smad3 specific antibody. The level of Smad2/3–Smad4 complex formation was determined by Western blot analysis with Smad4 antibody. The input lysate was probed against pSmad2, total Smad2, pSmad3, total Smad3, Smad4 and GAPDH. (D) A549 cells were transiently transfected with SBE4-Luc reporter plasmid and GLuc plasmid as internal control. Cells were treated with 10 µM SB431542, PP2, AG1478 and Lapatinib in presence of 10ng/ml TGFβ for 22h. Firefly luciferase activity was normalized to Gaussia luciferase and fold induction (over mock /DMSO treated) is expressed as the mean±SEM of triplicate measurements.