α tgn46 sheep polyclonal  (Bio-Rad)

Journal: bioRxiv

Article Title: MYO1C facilitates arrival at the Golgi apparatus through stabilization of branched actin

doi: 10.1101/409110

Figure Lengend Snippet: ( A ) Immunofluorescent staining of different Golgi marker, the matrix proteins GM130 (green), the cis -Golgi protein Giantin (red) and the trans -Golgi network protein TGN46 (magenta) in representative control (top) and MYO1C depleted (bottom) hTertRPE-1 cells in classical, unconstrained culture conditions. The nucleus is stained with Dapi (blue). Quantification of the 2D projected Golgi area in control conditions (n=145) and upon MYO1C depletion (n=165). Error bars represent the standard deviation of three independent experiments. *** indicates P-value < 1×10 −4 in a Student T-test. ( C ) Fluorescent images of representative hTertRPE-1 cells expressing GFP or GFP-MYO1C (green) and stained for the Golgi apparatus (red, GM130). ( D ) Quantification of the 2D projected Golgi area (stained by GM130) of non transfected cells and cells as in C. Error bars represent the standard deviation of three independent experiments with n > 40 cells of each condition. * indicates P-value < 1×10 −2 in a Student T-test on averages of three independent experiments ( E ) Electron microscopy images of representative intracellular areas of unconstrained hTertRPE-1 cells containing the Golgi apparatus in control conditions and upon MYO1C depletion. Scale bar: 1 µ m.

Article Snippet: We used the following mouse monoclonal antibodies α-GM130 (BD Biosciences cat: 610823), α-MYO1C (Santa Cruz sc-136544) and α-actin (Sigma, A-2228), rabbit polyclonal antibodies α-Lamp1 (Sigma, L1418 or Abcam, Ab24170), human monoclonal antibody (F2C-hFc) α-tubulin and α-Giantin (Recombinant Protein and Antibody Platform of the Institut Curie) and sheep polyclonal antibody α-TGN46 (Bio-Rad, AHP500).

Techniques: Staining, Marker, Standard Deviation, Expressing, Transfection, Electron Microscopy

Journal: bioRxiv

Article Title: MYO1C facilitates arrival at the Golgi apparatus through stabilization of branched actin

doi: 10.1101/409110

Figure Lengend Snippet: ( A ) Fluorescent images of a representative, unconstrained hTertRPE-1 cell stained with MYO1C antibody to visualize the endogenous protein (green), TGN46 antibody to visualize the Golgi apparatus (red) and Dapi to visualize the nucleus (blue). ( B, C ) Fluorescent images of representative, micropatterned hTertRPE-1 cells expressing GFP-MYO1C (green) and stained for the Golgi apparatus either with TGN46 or Giantin antibody (red) and the nucleus with Dapi (blue). ( D ) Fluorescent images of a time-lapse acquisition of unconstrained hTertRPE-1 cells expressing GFP-MYO1C (green) and mCherry-Rab6 (red). Scale bar: 10 µm.

Article Snippet: We used the following mouse monoclonal antibodies α-GM130 (BD Biosciences cat: 610823), α-MYO1C (Santa Cruz sc-136544) and α-actin (Sigma, A-2228), rabbit polyclonal antibodies α-Lamp1 (Sigma, L1418 or Abcam, Ab24170), human monoclonal antibody (F2C-hFc) α-tubulin and α-Giantin (Recombinant Protein and Antibody Platform of the Institut Curie) and sheep polyclonal antibody α-TGN46 (Bio-Rad, AHP500).

Techniques: Staining, Expressing

Journal: bioRxiv

Article Title: MYO1C facilitates arrival at the Golgi apparatus through stabilization of branched actin

doi: 10.1101/409110

Figure Lengend Snippet: ( A ) Fluorescent images of a representative, unconstrained hTertRPE-1 cell stained with MYO1C antibody to visualize the endogenous protein (green), TGN46 antibody to visualize the Golgi apparatus (magenta) and phalloidin to visualize the actin cytoskeleton (red). The nucleus is stained with Dapi (blue). ( B ) Fluorescent images of representative, micropatterned hTertRPE-1 cells expressing GFP-MYO1C (green) and stained with phalloidin to visualize F-actin (red) and Dapi to visualize the nucleus (blue). ( C ) Domain structure of MYO1C, the mutant form MYO1CΔABL and the truncated Tail domain. ( D ) Fluorescent images of representative hTertRPE-1 cells expressing GFP-MYO1C, GFP-MYO1CΔABL or GFP-Tail (green), stained for the Golgi apparatus (magenta, GM130) and F-actin with phalloidin (red). ( E ) Quantification of GFP-positive spots at the Golgi area as in D for GFP (n=24), GFP-MYO1C (n=49), GFP-MYO1CΔABL (n=56) or GFP-Tail (n=34). % of cells is shown that contain 0 (light grey), 1-10 (dark grey) or more than 10 (black) Golgi-associated spots per cell. Note that control (GFP-expressing) cells never contained more than 1 Golgi-associated spot per cell.

Article Snippet: We used the following mouse monoclonal antibodies α-GM130 (BD Biosciences cat: 610823), α-MYO1C (Santa Cruz sc-136544) and α-actin (Sigma, A-2228), rabbit polyclonal antibodies α-Lamp1 (Sigma, L1418 or Abcam, Ab24170), human monoclonal antibody (F2C-hFc) α-tubulin and α-Giantin (Recombinant Protein and Antibody Platform of the Institut Curie) and sheep polyclonal antibody α-TGN46 (Bio-Rad, AHP500).

Techniques: Staining, Expressing, Mutagenesis

Journal: The Journal of Cell Biology

Article Title: Optineurin links myosin VI to the Golgi complex and is involved in Golgi organization and exocytosis

doi: 10.1083/jcb.200501162

Figure Lengend Snippet: The loss of optineurin results in Golgi fragmentation. Mock (a–c) or optineurin siRNA-treated HeLa cells (d–f and d′–f′) were double labeled in immunofluorescence experiments with TGN46 (b, e, and e′) and GM130 (a, d, and d′). White boxes indicate areas enlarged in the pictures below (d′, e′, and f′). The Golgi fragments contain both marker proteins as indicated by the yellow color in the merged images (c, f, and f′). The arrows highlight the overlap between the marker proteins in (d′–f′). Bars, 10 μm. TEM analysis of mock (g) or siRNA-transfected (h) HeLa cells. Arrows indicate the position of Golgi stacks that can be found in both mock-transfected and siRNA-transfected cells. Bars, 200 nm.

Article Snippet: The following antibodies were used: affinity-purified rabbit pAb to human full-length optineurin (α-Optn; GenBank/EMBLDDBJ accession no. AF061034 ); monoclonal α-VSV-G to the luminal domain (a gift from R. Pepperkok and J. Simpson, EMBL, Heidelberg, Germany); polyclonal α-optineurin (Cayman); monoclonal α-GM130 (BD Transduction Laboratories); monoclonal α-TGN38 ( ); monoclonal α-GFP (Qbiogene); polyclonal α-GFP (Molecular Probes); monoclonal, polyclonal α-TGN46 (Serotec); and monoclonal and pAbs to the whole tail (α-MVI) and globular tail of myosin VI (α-MVI-GT; ; ).

Techniques: Labeling, Immunofluorescence, Marker, Transfection