Journal: The Journal of Biological Chemistry

Article Title: Slc35c2 Promotes Notch1 Fucosylation and Is Required for Optimal Notch Signaling in Mammalian Cells *

doi: 10.1074/jbc.M110.126003

Figure Lengend Snippet: Slc35c2 is primarily localized to Golgi in rat liver. A, a new polyclonal Ab against mouse and human Slc35c2 was tested on lysates of CHO cells expressing empty vector, mouse Slc35c1-Myc, mouse Slc35c2 with low transfection efficiency (C2(low)), or mouse Slc35c2-Myc and analyzed by Western blotting using anti-Slc35c2 C-terminal peptide antibody or anti-Myc. B, cell lysates from CHO cells transiently transfected with mouse Slc35c2 were treated with Endo H or PNGase F and analyzed by Western blotting using anti-Slc35c2 pAb. The membrane was stripped and re-probed with anti-Pofut1 C-terminal peptide pAb. Western blotting was performed using the Odyssey Infrared Imaging System. C, rat liver ER and Golgi fractions were assayed by immunoblotting using anti-Slc35c2 peptide or anti-PDI (ER marker) or anti-GM130 (Golgi marker) or anti-Pofut1 peptide antibodies. Western blotting was performed using the Odyssey Infrared Imaging System.

Article Snippet: Mouse anti-PDI mAb (1:400) and rabbit anti-PDI pAb (1:200) were obtained from Assay Designs (Ann Arbor, MI).

Techniques: Expressing, Plasmid Preparation, Transfection, Western Blot, Imaging, Marker

Journal: The Journal of Biological Chemistry

Article Title: Slc35c2 Promotes Notch1 Fucosylation and Is Required for Optimal Notch Signaling in Mammalian Cells *

doi: 10.1074/jbc.M110.126003

Figure Lengend Snippet: Slc35c2 subcellular localization in IEC and HeLa cells. Expression of Slc35c2 in fixed rat IEC (A–F) or HeLa cells (G–N). Transfected Slc35c2 was detected in B, H, J, and L with anti-Slc35c2 pAb; Slc35c2-Myc was detected in D, F, and N with anti-Myc mAb. Other antibodies included anti-PDI (ER marker), anti-ERGIC-53 (ERGIC marker), anti-GM130 (cis-Golgi marker), anti-ManII (medial-Golgi marker), and anti-β4GalT1 (trans-Golgi marker). Images were taken using a Leica SP2 AOBS confocal microscope. Scale bars, 10 μm.

Article Snippet: Mouse anti-PDI mAb (1:400) and rabbit anti-PDI pAb (1:200) were obtained from Assay Designs (Ann Arbor, MI).

Techniques: Expressing, Transfection, Marker, Microscopy