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. 2013 Nov 21;3:3288. doi: 10.1038/srep03288

Figure 3. AGO61 modifies GlcNAc residues at specific sites on α-DG.

Figure 3

(a) α-DG-Fc was transiently transfected with or without AGO61 into COS1 cells. α-DG-Fc recombinant proteins were collected from cell lysates and culture media using protein A resin and analyzed for laminin overlay and Western blot using anti-Fc and anti-O-GlcNAc antibodies. (b) α-DG373-HALO and its mutant T317A/T319A were transiently transfected with or without LARGE-myc into COS7 cells. HALO-fused proteins were collected from medium using HALO resin followed by digestion with TEV protease and then analyzed by laminin overlay. Cell lysates were analyzed for the expression of HALO-fused proteins and LARGE-myc by Western blot using anti-HALO and anti-myc antibodies. The full-length blots with anti-HALO and anti-myc antibodies are presented in Supplementary Figs. S7f and S7g, respectively. (c) α-DG373-HALO and its mutant T317A/T319A were transiently transfected with or without AOG61 into COS7 cells. HALO-fused proteins were collected from the cell lysates using HALO resin followed by digestion with TEV protease and then analyzed by Western blot using an anti-O-GlcNAc antibody (CTD110.6). The cell lysates were analyzed for the expression of HALO-fused proteins and AGO61 by Western blot using anti-HALO and anti-AGO61 antibodies. The full-length blots with anti-HALO and anti-AGO61 antibodies are presented in Supplementary Figs. S7h and S7i, respectively.