Alexa fluor 546 rabbit anti mouse igg

The following primary antibodies were used: rabbit anti-Pan-cadherin (C3678, Sigma-Aldrich, St. Louis, MO) that recognizes the conserved C-terminal domain of classic cadherins, mouse anti-N-cadherin (clone 32) and anti-E-cadherin (clone 36), both from BD Biosciences (Franklin Lakes, New Jersey, USA), rabbit anti-β-catenin (Invitrogen-Molecular Probes, Carlsbad, CA), mouse anti-active-β-catenin (clone 8E7, Millipore, Billerica, MA, USA), mouse anti-lamin A∖C (BD Biosciences), and mouse anti-α-tubulin (clone DM1a, Sigma-Aldrich). Secondary antibodies were Alexa Fluor™ 488 goat anti-rabbit IgG, Alexa Fluor™ 546 rabbit anti-mouse IgG (Invitrogen, Life Technologies, Brazil, São Paulo, SP, Brazil), and peroxidase-conjugated goat anti-rabbit and rabbit anti-mouse (Promega, Madison, WI). DAPI dihydrochloride (Invitrogen) was used for nuclear staining. The γ-secretase activity inhibitor Dapt (N-N[-(3,5-Difluorophenacetyl-l-alanyl)]-S-phenylglycine-t-butyl-ester) was from Merck Biosciences (Darmstadt, Germany). Nuclear and cytoplasmic fractions were extracted using NE-PER® Nuclear and Cytoplasmic Extraction Reagents (Pierce Biotechnology, Rockford, IL).