In experiments where antibodies were used to perturb E-cadherin-mediated cell–cell adhesions, MCF10A-E-cadherin-GFP cells were treated with 3 µg/ml of neutral, control antibody (76D5; gift of BM Gumbiner (Petrova et al., 2012 (link))), or E-cadherin-blocking antibody (DECMA-1; Abcam) for at least 2 hr prior to TFM measurements. As an additional control, cells were also imaged and measured pre-treatment.
In immunostaining experiments to visualize focal adhesions in MCF10A-E-cadherin-GFP cell clusters, a purified mouse anti-paxillin antibody (BD Biosciences, San Jose, CA) was used as the primary antibody and the Alexa Fluor 568 goat anti-mouse antibody (Life Technologies, Grand Island, NY) was used as the secondary antibody.
In immunostaining experiments to visualize focal adhesions in MCF10A-E-cadherin-GFP cell clusters, a purified mouse anti-paxillin antibody (BD Biosciences, San Jose, CA) was used as the primary antibody and the Alexa Fluor 568 goat anti-mouse antibody (Life Technologies, Grand Island, NY) was used as the secondary antibody.
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