Notch-Ras cooperation in breast carcinogenesis: (A) i. Figure represents soft agar colony formation assay of HMLE, HMLE-AcN1, HMLE-Raslow and HMLE-Raslow-AcN1 cells. ii. Graph shows tumor growth kinetics in nude mice for HMLE with (blue) & without (black) AcN1, and HMLE-Raslow cells with (red) & without AcN1 (green). iii. Graph represents soft agar colony formation of HMLE cells expressing various Ras effector loop mutants alone, or in combination with AcN1. HMLE-Rashigh cells were used as positive control. (B) Photomicrographs and bar graphs represent the effect of Notch activation (using DSL peptide), Notch inhibition (using GSI and DAPT) and inhibition of MAPK (using MEK1 inhibitor) on mammosphere formation; magnification 10×. (C) Photomicrographs represent immunostaining of cancer mammospheres for cleaved Notch1, Hes5 and phosphoErk1/2; magnification 40×. (D) Photomicrographs represent staining of normal and cancerous breast tissue for cleaved Notch1, Hes5 and phosphoErk1/2; magnification 20×. Insets represent higher magnification (40×) revealing nuclear reactivity.