Mammalian two-hybrid assays with the LRP1 or APP cytoplasmic domains. H4 human neuroglioma cells were transfected with the indicated constructs, and luciferase activity measured. A. The LRP1 cytoplasmic domain was cloned into the pBIND vector while the interactors JIP-1b, EB-1 and Nyggf4 were cloned into the pACT vector. Strong interactions between LRP1 and JIP-1b, EB-1 or Nyggf4 were observed, confirming results with yeast two hybrid studies. B. The APP cytoplasmic domain (known as APP intracellular domain/AICD) was cloned into pBIND, while the interactors JIP-1b, EB-1 and Nyggf4 were cloned into pACT. Strong interactions between JIP-1b or EB-1 with the APP intracellular domain were observed but no interaction was observed with Nyggf4 and the APP intracellular domain. ***, P < 0.0001.