Fig. 4

Interaction of DGCR2 with NRXN1. A DGCR2 interacts with NRXN1α in HEK 293T cells. Cells were co-transfected with FLAG-hDGCR2 and indicated constructs. Lysates were immunoprecipitated with anti-FLAG antibody and resulting complexes were blotted with anti-GFP and anti-FLAG antibodies. Inputs were blotted for perspective proteins as controls. B Surface DGCR2 interacts with NRXN1α in HEK 293T cells. Cells were co-transfected with FLAG-hDGCR2 and indicated constructs. Before lysis, the anti-FLAG antibody was added to the cells for intact cells IP. C ECD of DGCR2 interacts with NRXN1α in HEK 293T cells. Cells were co-transfected with NRXN1α-CFP and FLAG-hDGCR2, ΔICD, ΔECD or empty vector (Mock). Lysates were immunoprecipitated with anti-FLAG antibody and resulting complexes were blotted with anti-GFP and anti-FLAG antibodies. Inputs were blotted for perspective proteins as controls. D-E ECD of DGCR2 mediates its transcellular interaction with NRXN1 in HEK 293T cells. HEK 293T cells expressing NRXN1β or Mock along with RFP (red cells) were mixed with HEK 293T cells expressing NLGN1, Mock, FLAG-hDGCR2 or ΔECD along with GFP (green cells), respectively. An increasing amount of soluble ECD (0, 100 and 200 µL, respectively) was added to the cell mixtures of NRXN1β-expressing cells (red cells) and DGCR2-expressing cells (green cells). Cells were imaged, and aggregates were counted. Scale bar, 200 μm. From left to right in E, n = 12, 11, 10, 12, 9 or 10 images from three independent experiments. *** p < 0.001. n. s., p > 0.05, One-way ANOVA