Fig. 4. Inhibition of KRAS downstream signaling and 3D-cultured cell growth by inRas37 and BEZ-235 co-treatment. (A) MIA PaCa-2 and PANC-1 cells were treated with inRas37 (5 μM) and/or BEZ-235 (0.05 μM) for 6 h and then stimulated with EGF (200 ng/mL for MIA PaCa-2, and 50 ng/mL for PANC-1) for 10 min after serum starvation for 2 h. The expression levels of p-AKT, p-BRAF, and p-ERK were assessed using immunofluorescence staining. Scale bar, 30 μm. (B) Cell viabilities of MIA PaCa-2 and PANC-1 cells treated with inRas37 (5 μM) and/or BEZ-235 (0.05 μM) on every 2 days for 1 week were measured using the MTS assay. (C) Tumor spheroid assays were performed on MIA PaCa-2 and PANC-1 cells treated with inRas37 (5 μM), and/or BEZ-235 (0.05 μM) on every 2 days for 1 week, and sphere sizes were measured using Image J software. Scale bar, 200 μm. Data are presented as the means ± SD (***p<0.001).
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