A: Down-regulation of Hes-1 mRNA is seen in PDAC lines treated with MRK-003.
B: Colony formation in soft agar is inhibited in Pa03C cells with 5μM of MRK-003.
C: Normalized colony counts in PDAC cell lines, pre-treated with 2 and 5μM of MRK-003 (**, P<0.01). Three of the cells lines (Capan-1, Pa03C, and Pa14C) were “sensitive” based on significant inhibition of anchorage growth, while the other two (Pa16C and Pa29C) were “resistant”.
D and F: qRT-PCR and western blots of N1ICD stably transfected Pa03C showing fover-expression of transcripts corresponding to the ICD domain as compared to mock-trasfected cells.
F and G: Treatment of MRK-003 significantly reduced the number of colonies in mock Pa03C cells, while no significant effects are seen in N1ICD expressing Pa03C cells. Notably, expression of N1ICD per se results in a significant expansion of colonies in soft agar in untreated Pa03C compared to mock controls.