Figure 3 from Novel Syngeneic Cell Lines for Studying High-Risk BRAFV600E-Driven Colorectal Cancer In Vivo

Simultaneous inhibition of BRAFV600E and the HER family receptors leads to a reduction in signaling and viability in the NaJa cell lines. A, Representative Western blot (WB) analysis of NaJa cell lines, treated with 0.5 μmol/L encorafenib (E) and 1 μmol/L afatinib (A), either alone or in combination (E + A), for 24 hours. HSP90 and 14-3-3 serve as loading controls. B, Quantification of WB analysis of pEGFR and pERK normalized to the internal loading control (n = 3 biologically independent experiments). Data are presented as the mean ± SD. Statistical significance was calculated using one-way ANOVA and Dunnett multiple comparisons test for each cell line individually. C, Colony forming assays of NaJa cell lines treated for 10 days with encorafenib and afatinib, either alone or in combination. Shown is a representative result from three independent biological replicates. D, Quantification of the data shown in C. Data are presented as the mean ± SD and were normalized to the DMSO control (n = 3). Statistical significance was calculated using one-way ANOVA and Dunnett multiple comparisons test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. E, Representative surface plots illustrating the synergistic effects of encorafenib/afatinib cotreatment observed in XTT assays. F, Quantification of the synergy scores, as calculated by LOEWE (n = 4). A synergy score of >10 indicates a synergistic effect between two drugs. Data are presented as the mean ± SD.