Figure 4. Lnx1 interacts with... | Rockefeller University Press

Figure 4.

$Figure 4. Lnx1 interacts with postsynaptic EphB receptors. (A) The expression of membrane proteins in hippocampus from PW3 Lnx1−/− and WT mice was detected by Western blot and quantified. n = 3 mice per group. (B) Coimmunoprecipitation of Lnx1 and EphB1/2 in synaptosomes of hippocampus. Lnx1 was pulled down by EphB2 antibody from PSD fraction of either WT or EphB2K661R/K661R mice but neither EphB2−/− nor EphB2ΔVEV/ΔVEV mice. ns, nonspecific band; IB, immunoblot. (C) Schematic representation of p70-Lnx1 and its mutant constructs. (D) Immunoprecipitation (IP) of Lnx1 mutants and EphB1 or EphB2 coexpressed in NG108 cells. Lnx1 interacted with EphB1 through the N terminus and with EphB2 through the second PDZ domain. (E) Colocalization of Lnx1 and EphB2 (arrowheads in the right panels) in CA3 pyramidal cell layer (arrow in the left panel) were determined by immunostaining in PW3 WT mice. The higher-magnification views in white boxes indicate superresolution imaging of Lnx1 and EphB2. White dotted lines indicate neuronal boundary. Bars: 250 µm (left); 5 µm (bottom right); 0.5 µm (top right). Means ± SEM. Student’s t test (A); ***, P < 0.001.$

Lnx1 interacts with postsynaptic EphB receptors. (A) The expression of membrane proteins in hippocampus from PW3 Lnx1^−/− and WT mice was detected by Western blot and quantified. n = 3 mice per group. (B) Coimmunoprecipitation of Lnx1 and EphB1/2 in synaptosomes of hippocampus. Lnx1 was pulled down by EphB2 antibody from PSD fraction of either WT or EphB2^K661R/K661R mice but neither EphB2^−/− nor EphB2^ΔVEV/ΔVEV mice. ns, nonspecific band; IB, immunoblot. (C) Schematic representation of p70-Lnx1 and its mutant constructs. (D) Immunoprecipitation (IP) of Lnx1 mutants and EphB1 or EphB2 coexpressed in NG108 cells. Lnx1 interacted with EphB1 through the N terminus and with EphB2 through the second PDZ domain. (E) Colocalization of Lnx1 and EphB2 (arrowheads in the right panels) in CA3 pyramidal cell layer (arrow in the left panel) were determined by immunostaining in PW3 WT mice. The higher-magnification views in white boxes indicate superresolution imaging of Lnx1 and EphB2. White dotted lines indicate neuronal boundary. Bars: 250 µm (left); 5 µm (bottom right); 0.5 µm (top right). Means ± SEM. Student’s t test (A); ***, P < 0.001.