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. 2014 Dec 8;290(4):2137–2149. doi: 10.1074/jbc.M114.598821

FIGURE 5.

FIGURE 5.

EOGT mutants exhibit impaired O-GlcNAcylation of Notch1 EGF repeats. HEK293T cells were cotransfected to express N1EGF1–36-MycHis and EOGT constructs. A, N1EGF1–36-MycHis was immunoprecipitated (IP) from lysates using anti-Myc antibody and subjected to immunoblotting with CTD110.6 and anti-Myc antibodies. The specificity of the CTD110.6 antibody immunoreactivity was confirmed by a hapten inhibition test using 0.1 m GlcNAc. B, the levels of O-GlcNAc modification on N1EGF1–36-MycHis quantitated by densitometric analyses of immunoblots represented in A (n = 3; results are shown as mean ± S.D.; *, p < 0.01; **, p < 0.05; unpaired Student's t test). C, immunostaining of HEK293T transfectants with CTD110.6 (red), anti-calnexin (blue), and anti-Myc tag (cyan) antibodies. Bar, 10 μm.