Fig. 2.

Notch3 is a target of miR-221/222 in breast cancer cells. a MiR-221 and miR-222 expression levels were detected by qRT-PCR after forced transfection of miR-221 or miR-222 mimics in MCF-7 cells. b qRT-PCR of Notch3 mRNA after forced transfection of miR-221 and miR-222 mimics in MCF-7 cells. c Western blot analysis of endogenous Notch3, ERα, p27 and E-cadherin protein levels in MCF-7 cells after ectopic expression of miR-221/222 mimics in MCF-7 cells. d, e qRT-PCR of endogenous miR-221/222 and Notch3 mRNA in MDA-MB-231 cells after transfection with miR-221/222 inhibitors. f Western blot analysis of N3FL, N3ICD, ERα, p27, and Vimentin protein levels after transfection with miR-221/222 inhibitors in MDA-MB-231 cells. g Immunofluorescence staining of MCF-7 cells transiently transfected with GFP vector (expressing only GFP), GFP-miR-221 or GFP-miR-222 using an anti-Notch3 antibody. Notch3 signals (red) in miR-221 and miR-222-transfected cells are labeled with arrowheads. DAPI, 4′,6-diamidino-2-phenylindole