Expression of soluble WSX-1 enhances the binding of HN to F11 cells. F11 neurohybrid
2-Methoxyestradiol [s1, s2] (7 × 104 cells/well in 6-well plates), transfected with the backbone vector, WSX-1-FLAG-encoding vector or sWSX-1-FLAG-encoding vector, were used for HN immunofluorescence-based binding assays at 48 h after the start of transfection. The binding of HN to F11 cells was upregulated when WSX-1 was overexpressed in F11 cells [s2]. In a similar fashion, overexpression of sWSX-1 in F11 cells resulted in upregulation of the binding of HN to F11 cells. The binding manner between HN and F11
cells overexpressing sWSX-1 appeared to be nearly equal to that between HN and F11 cells overexpressing WSX-1 while the expression level of sWSX-1 was much higher than that of wild-type WSX-1 (Fig. 2D). This result suggests that sWSX-1 replaced full-length WSX-1 for the formation of the HN receptor and the endogenous expression level of CNTFR and gp130 may determine the stable binding capacity for HN in F11 cells.
