Fig. 4

Siglec-E interacts with CD36 independent of sialic acid binding activity. a HEK293 T cells were transfected with indicated vectors for 48 h. Cell lysates were prepared and subjected to immunoprecipitation with anti-Flag affinity resin. The expressions of Flag-CD36 and WT and mutant HA-SE protein in whole cell lysates (WCL) and immunoprecipitates were examined by Western blot analysis with indicated antibodies. b, c HEK293T cells overexpressing Flag-CD36 and WT HA-SE were pretreated with P-3FAX-Neu5Ac for 66 h or with 0.1 unit/ml of sialidase for 30 min at 37 °C prior to immunoprecipitation with anti-Flag affinity resin. Immunoprecipitates were then subjected to SNA lectin blotting (b) and Western blot analysis with indicated antibodies (c). d Peritoneal macrophages isolated from WT mice were treated without or with 0.1 unit/ml of sialidase for 30 min at 37 °C. Cleared cell lysates were subjected to immunoprecipitation with control IgG, anti-Siglec-E or anti-CD36 antibody as indicated. The immunoprecipitates were examined by Western blot analysis with indicated antibodies. The blots of immunoprecipitates were subjected to densitometry analysis and the relative quantitative results were shown under the blots. The value of control group for comparison in each blot was referred to 1