Figure 6.

TAK1 inhibitor synergizes with existing therapeutics to treat CLU deficient lung cancer. (A) NG25 synergizes with Trametinib to inhibit growth of H358 cells. 200 cells/well were seeded in 96 wells plate and cultured for 5 days with indicated treatment. Viability of cells analyzed with CCK8 assay. Statistics done on day 5 with two-tailed t-test. (B) NG25 synergizes with Trametinib to inhibit 2-D colony formation of H358 cells. Cells were culture for 7 days with indicated treatment. (C) Quantification of B, one-way ANOVA test. (D) NG25 synergizes with Trametinib to inhibit growth of Hop62-shCLU derived xenograft tumor. Hop62-shCLU cells (3 million) were s.c. implanted in the flanks of nude mice. 1-week post implantation, mice were treated with NG25 (4 mg/kg/Day, intravenous injection), Trametinib (Tram, 1 mg/kg/Day, gavage), or combination for 15 days. n > 4 in each group. two-tailed t-test. (E) Image of tumors harvested in D. (F) Weight of tumors harvested in D. one-way ANOVA test. (G) NG25 synergizes with Trametinib to shrink CLU deficient Kras^G12D driven lung cancer in transgenic mouse model. Lsl-Kras^G12D/+ mice were treated with pSECC-sgCLU lentivirus by nasal inhalation. Tumor burdens were documented with CT. Mice were treated as indicated. NG25 (4 mg/kg/Day, intravenous injection), Trametinib (Tram, 1 mg/kg/Day, gavage), or combination for 12 days. n > 4 in each group. (H) relative tumor volume of mice of G. (I) Representative pathology images of mice of G. (J) Quantification of tumor numbers of mice of G. (K) Quantification of size of tumor in G. one-way ANOVA test on comparison of singlet treatment with the combo treatment. All the data plotted are mean ± s.e.m., ***P < 0.005, ****P < 0.0001, n = 3