Association of the zonula adherens with mRNAs. Homo sapiens

We report the association of the zonula adherens with a set of mRNAs, through the E-cadherin-p120 catenin binding partner called PLEKHA7. Overall design: Isolation and identification of mRNAs associated to the zonula adherens in Caco2 colon cells, by formaldehyde intracellular cross-linking, RNA-immunoprecipiation (RNA-IP) by using a PLEKHA7 antibody (Sigma, HPA038610) and subsequent RNA isolation and RNA-sequencing. An IgG immunoglobulin was used in parallel and on the same lysates as negative RNA-IP control to exclude non-specific mRNA bindning, whereas total Caco2 mRNA was also isolated from the same cells and subjected to RNA sequencing, in order to caclulate fold-enrichment of mRNAs in the PLEKHA7 RNA-IP compared to total cellular mRNA content. Four indepedent experiements were performed.