Supplementary Figures 1 - 19 from Failure to Induce Apoptosis via BCL-2 Family Proteins Underlies Lack of Efficacy of Combined MEK and PI3K Inhibitors for KRAS-Mutant Lung Cancers

PDF file - 11671KB, Supplemental Figure 1. Response of Kras mouse models to MEKi/PI3Ki. Supplemental Figure 2. MEKi/PI3Ki induces G1 arrest and inhibits proliferation of KRAS mutant NSCLC cell lines. Supplemental Figure 3. Combined MEK and PI3K inhibition is necessary for maximal reduction in cell proliferation. Supplemental Figure 4. Apoptotic response of KRAS mutant NSCLC cell lines in response to MEKi/PI3Ki. Supplemental Figure 5. Mutational status of TP53 or STK11/LKB1 does not correlate with apoptotic response to MEKi/PI3Ki. Supplemental Figure 6. Secreted Gaussia luciferase allows for precise quantitation of tumor growth and treatment response. Supplemental Figure 7. Tumor response of KRAS NSCLC xenografts to MEKi/PI3Ki. Supplemental Figure 8. Apoptosis induced by MEKi/PI3Ki is BAX and caspase-3 dependent. Supplemental Figure 9. Modulation of MEK/ERK and PI3Ki/AKT transcriptional output does not correlate with apoptotic sensitivity of KRAS mutant NSCLC cancer cells. Supplemental Figure 10. Modulation of RalGDS signaling does not correlate with apoptotic sensitivity of KRAS mutant NSCLC cell lines. Supplemental Figure 11. Sensitivity to MEKi/PI3Ki does not correlate with BH3 priming. Supplemental Figure 12. siRNA mediated knockdown of PUMA and BIM protects from apoptosis induced by MEKi/PI3Ki. Supplemental Figure 13. Inhibition of BCL-2 alone does not restore apoptotic response in insensitive KRAS mutant NSCLC cell lines. Supplemental Figure 14. Individual protein expression levels of BCL-2 family members do not correlate with sensitivity to MEKi/PI3Ki. Supplemental Figure 15. Inducible ectopic expression of BIM. Supplemental Figure 16. Restoration of apoptosis by ABT-263 leads to MEKi/PI3Ki-induced regression in vivo. Supplemental Figure 17. In vitro derived MEKi/PI3Ki resistant cells. Supplemental Figure 18. Cell lines derived from resistant Kras p53L/L tumors. Supplemental Figure 19. PUMA and BIM mRNA levels do not differ between cell lines derived from treatment naive and resistant Kras p53L/L tumors. Supplemental Table 1. Mutational status of KRAS mutant NSCLC cell lines. Supplemental Table 2. Knockdown efficiency of pLKO shRNA hairpins used in lentiviral screen.