We are working with HUVECS that are adhesive cells, but according to my boss they should be able to form AJs while in suspension, so that shouldn't be an issue. I guess the biggest challenge for us would be establishing the experimental setup itself. I still haven't had time to read your protocol in detail, but I already noticed it is fairly complex. Would you be available for helping us with some technical details or doubts we may have if we decide to give it a try? Thank you so much for all the help!

Hey Christine! Actually I am designing primers for human, and yes, I am using Sybr Green. I have designed a couple of pairs already using different genomic regions with potential binding sites (both in the promoter and in the first intron of the gene) and also others in neighbouring regions that do not contain any binding sites to use as internal controls. I also did a set of primers in a different unrelated gene that has already been proved to interact with the transcription factor I am interested in and that I will be using as a positive control. Now all that is left to do is order and test them! Thank you so much for these tips, they were really useful! Good luck for your experiments as well! Joana