Rabbit e cadherin antibody

Two-cell embryos were collected at day 2 of pregnancy and cultured in single-step medium for another 2-day period. Hatched blastocysts were added to the confluent monolayers of OPN-targeted siRNA or NC-siRNA pre-treated mESC in DMEM/F12 containing 10% cFBS for 48 h. Immunofluorescence was performed on this embryo-mESC co-culture as previously described [29] (link). Cells were incubated with a rabbit E-cadherin antibody (1∶100 dilution, Cell Signaling Technology, Boston, USA), goat polyclonal MMP-9 antibody (1∶100 dilution, Santa Cruz) or rabbit Ig G (1∶100 dilution, Santa Cruz), followed by the FITC conjugated secondary antibody (PIERCE, Rockford, IL, USA). Nuclei were stained with DAPI (Zhongshan Golden Bridge Bio-technology, Beijing, China). The trophoblast spreading area was quantified by measuring the E-cadherin signal area using the Image J software. A total of 10 embryos were calculated in each experiment, and the experiments were repeated three times.

To detect actin, E-cadherin and vimentin in MCF10A, cells were fixed with 4% paraformaldehyde for 15min and permeabilized with 0.1% Triton X-100 in PBS before incubation with rabbit E-cadherin antibody (Cell Signaling) followed by alexa 555-conjugated anti-rabbit (Cell Signaling), vimentin antibody (Dako) followed by alexa 555-conjugated anti-mouse (Cell Signaling) or rhodamine-phalloïdin (Interchim, Montluçon, France). The slides were washed, mounted and viewed using an automated LEICA DMRXA2 microscope.