Fig. 8

Pharmacologic targeting of NOTCH signaling in triple-negative breast cancer (TNBC) cells. a Immunofluorescence analysis showing representative images of MDA-MB-231 and MDA-MB-231 lung metastasis (LM) TNBC cells stained in green with a NOTCH3 polyclonal antibody. Nuclei are stained in blue with 4′,6-diamidino-2-phenylindole (DAPI). Graph shows the average number of NOTCH3-expressing cells from three independent experiments (± SD). b In vitro real-time invasion assay of MDA-MB-231 LM TNBC cells infected with scramble lentivirus short hairpin RNAs (lenti-shRNAs; control) and lenti-shRNAs targeting NOTCH3 messenger RNA. Graph shows the average number of invasive cells from three independent experiments (± SD). c Experimental lung metastasis imaging in live animals of LY-411575-treated or dimethyl sulfoxide (DMSO)-treated MDA-MB-231 LM cells expressing the firefly luciferase reporter lentivector after tail vein injection. d Lungs isolated from nude mice that were injected with LY-411575-treated or DMSO-treated MDA-MB-231 LM TNBC cells. Following 4 weeks of growth, animals were killed, and lung tissues were stained with H&E to determine the presence of metastatic lesions as previously described [29]