Figure 4

Alteration of signal transduction by Env and Sprouty2. (A) Cell lysates were prepared by homogenization of A549, A549-Spr, A549-Env, BEAS-2B and BEAS-2B-Env cells. The proteins were separated on 10% acrylamide gel, transferred to nitrocellulose membrane and probed with various antibodies. The results show the levels of phospho p38 MAPK, p38 MAPK, phospho p44/42 ERK, p44/42 ERK, phospho Ser473 Akt, Akt, TWIST, phospho PTEN, PTEN and β-actin in the indicated cell lines. (B) Proteins were extracted from cytoplasmic (cy) and nuclear (nu) fractions of the cells and the levels of phospho STAT3 and STAT3 were ascertained by Western blot. (C) MMP Zymogram: Conditioned media obtained from the cell lines was assayed for gelatinase activity using 10% SDS-PAGE gel to estimate MMP activity. (D) Conditioned media obtained from the cell lines was concentrated; the secreted proteins were separated by SDS-PAGE and immunoblotted with antibodies against TIMP1 and TIMP2.