Fig. 7.

Co-expression of Cbln1 with Cbln4 alters the receptor binding specificity of HA-Cbln4. HEK293T cells were transfected with DCC, Nrxn3β or empty vector (Mock). Cells were subsequently exposed to conditioned medium (Input) derived from HEK293 cells co-transfected with either HA-Cbln4 and mock plasmid or HA-Cbln4 and Cbln1 for 4 h. After washing with medium, cells were lysed with sample buffer and subjected to SDS-PAGE and immunoblotted with anti-hemagglutinin (HA), anti-Cbln1 or anti-Flag tag antisera. HA-Cbln4 shows strong binding to DCC and minimal binding to Nrxn3β. Co-expression of Cbln1 with HA-Cbln4 alters the binding specificity of HA-Cbln4 by reducing its association with DCC and promoting its association with Nrxn3β.