Description

Co-expression of wild-type Arf1-GFP (Arf1(WT)-GFP) (gray) and inactive Src (SrcK-DsRed) (red) in Src-deficient SYF fibroblasts does not induce trafficking of Helix Pomatia Lectin (HPL) (green) from the Golgi. HPL binds various glycans but the Tn antigen in particular. The Tn antigen refers to terminal α-linked N-acetyl galactosamine residues (GalNAc) linked to Ser or Thr residues. HPL indicates distribution of GalNac-Ts. Inactive Src is highly enriched at the Golgi. Cells were fixed for 10 min (4% paraformaldehyde) and permeabilized (0.2% Triton X-100) prior to staining with Hoechst (blue) and Alexa 647-conjugated-HPL. Cells were mounted onto glass slides using FluorSave (Merck) and imaged at room temperature using an inverted FluoView confocal microscope (model IX81; Olympus) with either a 60x objective (U Plan Super Apochromatic, UPLSAPO; NA 1.35) or 100x objective (UPLSAPO; NA 1.40) using Immersol oil. Microscope coupled with a CCD camera (model FVII). Images were acquired and processed using Olympus FV10-ASW software. Image corresponds to Fig 7F in J Cell Biol. 189: 843-858. 2010. Images in Fig 7 include CIL#s 13571, 13572, 13573, 13574, 13575, 13576, 13577.