Figure 1

Confirmation of the interaction between HcESPs and goat T cells and identification of the immune complexes from the interacting proteins by co-IP assays. A Collection of HcESPs and generation of rat anti-HcESP IgG. Lane 1: SDS-PAGE analysis of HcESPs; lane 2: Immunoblot analysis of HcESPs using rat anti-HcESP IgG as the primary antibody; lane 3: Immunoblot analysis of HcESPs using normal rat IgG as the negative control. B Goat T cell sorting by MACS. The purity of the isolated T cells was above 95%, as indicated by flow cytometry analysis. C Binding of HcESPs to goat T cells in vitro. The immunocytochemistry assays were performed using rat anti-HcESP IgG or normal rat IgG (control). DAPI (blue) and Cy3-conjugated secondary antibodies (red) were utilized for dual staining. T cells stimulated in the absence (a) or presence (b) of HcESPs were incubated with rat anti-HcESP IgG as the primary antibody. T cells pretreated with HcESPs were incubated with normal rat IgG as the primary antibody (c). Scale bars, 50 µm. D The interaction between HcESPs and T cells was tested by co-IP assays. Lane 1: SDS-PAGE analysis of the cell lysates precipitated by rat anti-HcESP IgG; lane 2: SDS-PAGE analysis of the cell lysates precipitated by normal rat IgG (control); lane 3: Western blot analysis of the cell lysates precipitated by rat anti-HcESP IgG; lane 4: Western blot analysis of the cell lysates precipitated by normal rat IgG (control); lane M: standard protein molecular marker.