Staining was conducted as previously published [11 (link), 22 (link)]. RFP staining: rabbit anti-RFP (Abcam, Cambridge, UK) 1:30 dilution, Alexa Fluor® 488 goat anti-rabbit secondary (Invitrogen); CDH1 staining: rabbit E-cadherin antibody (24E10; Cell Signaling Technology, Boston, MA, USA) 1:400 dilution. Internal negative controls were exposed to rabbit IgG or 10% goat serum rather than primary antibody. Images were captured on a Nikon TE2000 inverted microscope with IPLab software (BD Biosciences, Rockville, MD, USA), original magnification at 200× (RPF) or 400× (CDH1).
Metastases were calculated as a ratio of the number of RFP-positive cells versus the total amount of cells (determined by DAPI nuclear stain) per field of view. Points represent the ratio of RPF-positive cells versus DAPI for each section examined with mean for each group represented as horizontal black bar. SEM for each group indicated in red.
Metastases were calculated as a ratio of the number of RFP-positive cells versus the total amount of cells (determined by DAPI nuclear stain) per field of view. Points represent the ratio of RPF-positive cells versus DAPI for each section examined with mean for each group represented as horizontal black bar. SEM for each group indicated in red.
