Western blot analysis of VISTA from cell lysates revealed multiple bands ranging from 40 to 70 kDa, indicating glycosylation of VISTA; treatment with peptide-N-glycosidase F (PNGase F), which removes global N-linked glycosylation products, dramatically decreased the molecular weight of the VISTA protein with a homogeneous pattern of ~36 kDa in CRC cell lines (Fig. 6D). Here, VSIR is linked to colorectal carcinoma.