Knockdown of c-Met and E-cadherin in OKF6/TERT2 cells

As previously described [51 (link)], OKF6/TERT2 cells were grown in 6 well plates to 80% confluency and transfected with 40 pmole of c-Met siRNA (#SC-29397, Santa Cruz Biotechnology) or E-cadherin siRNA (#SC-35242, Santa Cruz Biotechnology) using Lipofectamine 2000 (#11668027, ThermoFisher Scientific) following the manufacturer’s instructions. After 24 h, the cells were trypsinized, seeded onto fibronectin coated glass coverslips, and incubated for another 24 h before use. The extent of protein knockdown was determined by immunoblotting with an anti-c-Met antibody (#8198, Cell Signaling Technology), anti-E-cadherin antibody (#3195, Cell Signaling Technology), anti-EGFR antibody (#4267, Cell Signaling technology), and anti-β-actin antibody (#A5441, Sigma-Aldrich).

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Phan Q.T., Solis N.V., Cravener M.V., Swidergall M., Lin J., Huang M.Y., Liu H., Singh S., Ibrahim A.S., Mazzone M., Mitchell A.P, & Filler S.G. (2023). Candida albicans stimulates formation of a multi-receptor complex that mediates epithelial cell invasion during oropharyngeal infection. PLOS Pathogens, 19(8), e1011579.

Publication 2023

c-Met siRNA E-cadherin siRNA Lipofectamine 2000 anti-c-Met antibody anti-E-cadherin antibody anti-EGFR antibody anti-β-actin antibody

Actin Anti antibody Anti c antibody Cadherin Cells E cadherin Egfr Fibronectin Lipofectamine 2000 Protein Sirna

Corresponding Organization : VIB-KU Leuven Center for Cancer Biology

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Variable analysis

independent variables

C-Met siRNA (#SC-29397, Santa Cruz Biotechnology)
E-cadherin siRNA (#SC-35242, Santa Cruz Biotechnology)

dependent variables

Extent of protein knockdown determined by immunoblotting with anti-c-Met antibody (#8198, Cell Signaling Technology)
Extent of protein knockdown determined by immunoblotting with anti-E-cadherin antibody (#3195, Cell Signaling Technology)
Extent of protein knockdown determined by immunoblotting with anti-EGFR antibody (#4267, Cell Signaling technology)
Extent of protein knockdown determined by immunoblotting with anti-β-actin antibody (#A5441, Sigma-Aldrich)

control variables

OKF6/TERT2 cells grown in 6 well plates to 80% confluency
Lipofectamine 2000 (#11668027, ThermoFisher Scientific) used for transfection according to manufacturer's instructions
Cells trypsinized, seeded onto fibronectin coated glass coverslips, and incubated for 24 h before use

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