Figure 2 from Novel Syngeneic Cell Lines for Studying High-Risk BRAFV600E-Driven Colorectal Cancer In Vivo

NaJa cells differ in their response to drug treatment. A, Colony formation assay of NaJa cells treated with vemurafenib (V), encorafenib (E), and trametinib (T) at the indicated concentrations for 10 days. Shown is a representative result from three biological replicates. B–D, Quantification of the data shown in A. 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 < 0.0001. E, Phospho-RTK array of NaJa cells treated with 1 μmol/L vemurafenib (Vem), 0.5 μmol/L encorafenib (Enco), or 50 nmol/L trametinib (Tram) for 5 days. Shown are the log2 FCs of treated cells versus DMSO control. Array membranes and cell line specific heatmaps are displayed in Supplementary Fig. S7. F, Western blot showing the effect of a 5-day treatment of the NaJa cells on various signaling pathways. G, Western blot and quantification of HER2 and HER3 (n = 4). Basal EGFR expression was quantified based on data in (F), and five additional biological replicates normalized to the internal loading control. Data are presented as the mean ± SD and were normalized to NaJa-F for comparison. Statistical significance was calculated using one-way ANOVA and Tukey multiple comparisons test. *, P < 0.05; **, P < 0.01; ****, P < 0.0001. H, Bulk RNA-seq of NaJa cells treated with 0.5 μmol/L encorafenib for 1 day or 5 days showing upregulation (red) or downregulation (blue) of ERK target genes and the adjusted P value (bubble size).