Fig. 2
Anti-proliferative activity of AZ304 and Cetuximab in CRC cell lines with different BRAF mutation status. Two V600E mutant BRAF cell lines (RKO, HT-29) and two wild-type BRAF cell lines (DiFi, Caco-2) were treated with DMSO or increasing concentrations of AZ304 (0, 0.1, 1, 10, 100 μM), for 48 h (a) and 72 h (b). Viable cells were determined by MTT assay. c EdU incorporation assay. Four cells lines from (a) were treated with DMSO or indicated AZ304 for 24 h, followed by incubation with EdU and Hoechst in sequence. Hoechst 33342 (blue) and EdU (red) represent cell nuclei and nuclei of proliferative cells, respectively. The percentages of the EdU-positive cells are presented (right). Student’s t-tests were used for statistical analyses. Data are plotted as mean ± SD. *P < 0.05 vs. control; **P < 0.01 vs. control. d These four cells lines from (a) were treated with DMSO or 2 uM AZ304 for 6 h. Expression levels of phosphorylation ERK were analysed by Western blot. e BRAF mutant cell line RKO, and BRAF wild type cell line Caco-2 were treated with DMSO or 2 uM AZ304 for the indicated times. Expression levels of phosphorylation ERK were analysed by Western blot. The four cell lines were treated with increasing concentrations of Cetuximab (0, 0.1, 1, 10, 100 μg/ml) for 48 h (f) and 72 h (g). Cell viability was determined by MTT assay
