Figure 7. TGF-βR Inhibitor and TKIs Synergize to Suppress Proliferation of MED12^KDNSCLC Cells.

(A) Combination of TGF-βR and ALK inhibitors synergistically inhibits growth of MED12^KD NSCLC cells harboring EML4-ALK translocation. H3122 cells expressing pRS or shMED12 vectors were cultured in the absence and the presence of 1 μM LY2157299, 300 nM crizotinib, or the combination of 1 μM LY2157299 and 300 nM crizotinib. The cells were fixed, stained, and photographed after 14 (untreated and LY2157299 alone) or 28 days (crizotinib alone and LY2157299 plus crizotinib).

(B) Combination of TGF-βR and EGFR inhibitors synergistically inhibits growth of MED12^KD NSCLC cells harboring EGFR-activating mutation. PC9 cells expressing pLKO or shMED12 vectors were cultured in the absence and the presence of 1 μM LY2157299, 100 nM gefitinib, or the combination of 1 μM LY2157299 and 100 nM gefitinib. The cells were fixed, stained, and photographed after 10 (untreated and LY2157299 alone) or 28 days (gefitinib alone and LY2157299 plus gefitinib).

(C and D) Combination of LY2157299 with crizotinib or gefitinib suppressed the ERK activation driven by MED12^KD in both H3122 and PC9 cells. (C) H3122 cells were grown in the absence or presence of 20 μM NVP-TAE684, 5 μM LY2157299, or the combination of 20 μM NVP-TAE684 and 5 μM LY2157299 for 6 hr, and the cell lysates were harvested for western blotting analysis. (D) PC9 cells were grown in the absence or presence of 25 nM gefitinib, 5 μM LY2157299, or the combination of 25 nM gefitinib and 5 μM LY2157299 for 6 hr, and the cell lysates were harvested for western blotting analysis.

See also Figure S4.