Figure 3. Inhibition of TRAF6 impairs growth and tumor formation of lung cancer cells.

(A) Growth curves of cell lines with TRAF6 inhibition (shT6) is shown relative to vector-expressing (pLKO.1 control) cells. (B and C) Soft agar colony formation for the indicated cell lines after TRAF6 inhibition. Original magnification, ×20. (D) Cell lines H460 and HCC95 were transduced with virus encoding shT6. After TRAF6 inhibition, cell survival was evaluated by flow cytometry for Annexin V expression. HCC95 cells transduced with vector or shT6 were also cultured in liquid media, and morphological assessment confirmed that shT6-expressing cells exhibited features of increased cell death, such as cell detachment and shrinkage and membrane blebbing. Original magnification, ×40. (E–G) Cell lines H1395 (E), H460 (F), and HCC95 (G), transduced with vector (pLKO.1-control) or shT6, were subcutaneously injected into NOD/SCID mice. Tumor volume is shown at the experimental endpoint. *P < 0.05.