Fig. 5

The CD146/Galectin-9 axis promotes pRBC and lymphocyte adhesion to BBBECs. a Silver staining of proteins from immunoprecipitation samples (n = 3). b Western blot analysis of Gal-9 expression in the EL4 and bEnd.3 cell lines. Actin served as a loading control. c FACS analysis of membrane-bound Gal-9 on primary BBBECs, T cells, and RBCs from healthy and eCM mice. d Immunoprecipitation analysis of the interaction between CD146 and Gal-9 in bEnd.3 and T cells. e Pull-down analysis using sCD146 and Gal-9-FC proteins. f Fc or Fc-Gal-9 was added to wells coated with different concentrations of CD146, and ELISA was performed. g Pull-down analysis of CD146 and Gal-9 in the presence of the anti-CD146 antibody AA98 or control mIgG antibody. h Adhesion assay of T cells with knockdown or overexpression of Gal-9 and bEnd.3 cells with knockdown or overexpression of CD146 (n = 3). i Adhesion assay of pRBCs isolated from Gal-9-WT or Gal-9-KO eCM mice and primary BBBECs isolated from CD146-WT or CD146-KO mice. *p < 0.05, **p < 0.01, ***p < 0.001. The data are representative of at least three independent experiments