Figure 8.

Modification of SDC1 core proteins with glycosaminoglycan chains and the interaction between N-cadherin and SDC1. A, gene expression levels of SDC1, SDC2, SDC3, and SDC4 in BT-549 (open boxes) and C4ST-1KO cells (shaded boxes) were analyzed by real-time PCR (n = 3). Statistical significance was assessed using a Student's t test. B, after the cell lysates of BT-549 and C4ST-1KO cells were digested with heparitinase and heparinase (HSase&Hepase) and/or chondroitinase ABC (CSase ABC) as indicated, the digests were subjected to immunoblotting using anti-SDC1 (Prestige Antibodies®, catalog no. HPA006185, Sigma) or anti-actin antibodies. C, BT-549 and C4ST-1KO cells were immunostained with anti-SDC1 and anti-N-cadherin antibodies. Magnified images of the areas within white boxes 1 (panel c, BT-549) and 2 (panel g, C4ST-1KO) are shown in panels d and h, respectively. D, cell lysates digested with heparitinase, heparinase, and chondroitinase ABC were immunoprecipitated in the presence or absence (w/o antibody) of anti-N-cadherin (IP, anti-N-cadherin antibody (Ab)) or anti-SDC1 antibodies (IP, anti-SDC1 antibody). Immunoprecipitates were analyzed by immunoblotting using the indicated antibodies.