rabbit polyclonal anti dync1h1  (Bethyl)

Journal: eLife

Article Title: Identification of host proteins differentially associated with HIV-1 RNA splice variants

doi: 10.7554/eLife.62470

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-DYNC1H1 (rabbit polyclonal) , ThermoFisher , PA5-49451; RRID: AB_2634905 , IF (1:200) WB (1:1000).

Techniques: Stable Transfection, Expressing, Recombinant, Plasmid Preparation, Sequencing, Transfection, Construct, Knockdown, Reverse Transcription, Extraction, Transferring, Hybridization, Software

Journal: Neural Development

Article Title: Analysis of a zebrafish dync1h1 mutant reveals multiple functions for cytoplasmic dynein 1 during retinal photoreceptor development

doi: 10.1186/1749-8104-5-12

Figure Lengend Snippet: The cnb locus encodes dync1h1 . (A) Map of the cnb critical genomic region and exon/intron representation of dync1h1 . Linked microsatellite markers and recombinants are placed onto the ZV7 zebrafish genomic map. (B) RT-PCR results from two independent cDNA samples each of cnb or wild-type embryos. Top image of bands from reactions with dync1h1 primers and bottom image of bands from reactions with EF1alpha primers. (C) Chromatograms showing wild-type (top) and cnb sequence flanking the C-to-A mutation. (D) Schematic of the Dync1h1 protein and location of the nonsense mutation. (E) Location of retrovirus insertion for dync1h1 mutant allele hi3684.

Article Snippet: Antibodies included: mouse monoclonal anti-acetylated-α-tubulin (Sigma-Aldrich, St Louis, MO, USA); rabbit polyclonal anti-human Dync1h1 (amino-terminal 321 amino acids; serum #46, gift from Dr Richard Vallee, Columbia University); rabbit polyclonal anti-human Dync1h1 (carboxy-terminal 400 amino acids; 12345-1, ProteinTech, Chicago, IL, USA); rabbit polyclonal anti-rat Dync1h1 (amino acids 4,320 to 4,644; Dynein HC, R-325: sc-9115, Santa Cruz Biotechnology, Santa Cruz, CA, USA); rabbit polyclonal anti-Dync1i1 (gift from Dr Richard Vallee, Columbia University); rabbit polyclonal anti-Dynct1/p150 antibody (gift from Dr Kevin Vaughan, University of Notre Dame); mouse monoclonal anti-SV2 (Developmental Studies Hybridoma Bank).

Techniques: Reverse Transcription Polymerase Chain Reaction, Sequencing, Mutagenesis

Journal: Neural Development

Article Title: Analysis of a zebrafish dync1h1 mutant reveals multiple functions for cytoplasmic dynein 1 during retinal photoreceptor development

doi: 10.1186/1749-8104-5-12

Figure Lengend Snippet: Morpholino knock-down of dync1h1 phenocopies cnb mutants . (A, B) Dorsal views and retinal histology of control (cntl) morpholino (MO) injected (A) and dync1h1 ATG MO injected (B) embryos shown at 3.5 dpf. Note the smaller eyes, expanded melanophores, and photoreceptor dysmorphogenesis. (C) Western blot of wild-type (WT) versus cnb embryos at 3, 4, and 5 dpf (left blot) and wild-type and dync1h1 morphants at 2 dpf (right blot). Each blot was cut and probed with either antisera against human DYNC1H1 (amino-terminal 321 amino acids; top) or gammaTubulin (bottom). Scale bar: 40 μm for retinal histology.

Article Snippet: Antibodies included: mouse monoclonal anti-acetylated-α-tubulin (Sigma-Aldrich, St Louis, MO, USA); rabbit polyclonal anti-human Dync1h1 (amino-terminal 321 amino acids; serum #46, gift from Dr Richard Vallee, Columbia University); rabbit polyclonal anti-human Dync1h1 (carboxy-terminal 400 amino acids; 12345-1, ProteinTech, Chicago, IL, USA); rabbit polyclonal anti-rat Dync1h1 (amino acids 4,320 to 4,644; Dynein HC, R-325: sc-9115, Santa Cruz Biotechnology, Santa Cruz, CA, USA); rabbit polyclonal anti-Dync1i1 (gift from Dr Richard Vallee, Columbia University); rabbit polyclonal anti-Dynct1/p150 antibody (gift from Dr Kevin Vaughan, University of Notre Dame); mouse monoclonal anti-SV2 (Developmental Studies Hybridoma Bank).

Techniques: Knockdown, Control, Injection, Western Blot

Journal: Neural Development

Article Title: Analysis of a zebrafish dync1h1 mutant reveals multiple functions for cytoplasmic dynein 1 during retinal photoreceptor development

doi: 10.1186/1749-8104-5-12

Figure Lengend Snippet: Photoreceptors from cnb / dync1h1 mutants show cell-autonomous defects in morphogenes . (A-F) Images of histology sections from genetically mosaic retinas of wild-type (wt) donor, wild-type host (A-C) or cnb donor, wild-type host (D-F). Two examples are shown at low magnification to indicate overall clone size and one example at higher magnification to show photoreceptor morphology. Donor cells are label with the brown lineage tracer. Black retinal pigment epithelium cells provide reference for orientation. Note the lack of elongated morphology in cnb photoreceptor cells. Scale bars: 40 μm in (A, B, D, E); 15 μm in (C, F). (G) Schematic showing how nuclear position was evaluated. Dashed lines represent the external limiting membrane (upper) and apical side of the outer plexiform layer (bottom) for the host retina. (H) Scatter plot of nuclear position (c/a ratio) for donor cells of wild-type (black, left side) or cnb (grey, right side) genotypes.

Article Snippet: Antibodies included: mouse monoclonal anti-acetylated-α-tubulin (Sigma-Aldrich, St Louis, MO, USA); rabbit polyclonal anti-human Dync1h1 (amino-terminal 321 amino acids; serum #46, gift from Dr Richard Vallee, Columbia University); rabbit polyclonal anti-human Dync1h1 (carboxy-terminal 400 amino acids; 12345-1, ProteinTech, Chicago, IL, USA); rabbit polyclonal anti-rat Dync1h1 (amino acids 4,320 to 4,644; Dynein HC, R-325: sc-9115, Santa Cruz Biotechnology, Santa Cruz, CA, USA); rabbit polyclonal anti-Dync1i1 (gift from Dr Richard Vallee, Columbia University); rabbit polyclonal anti-Dynct1/p150 antibody (gift from Dr Kevin Vaughan, University of Notre Dame); mouse monoclonal anti-SV2 (Developmental Studies Hybridoma Bank).

Techniques: Membrane

Journal: Neural Development

Article Title: Analysis of a zebrafish dync1h1 mutant reveals multiple functions for cytoplasmic dynein 1 during retinal photoreceptor development

doi: 10.1186/1749-8104-5-12

Figure Lengend Snippet: Specimens analyzed for ultrastructure

Article Snippet: Antibodies included: mouse monoclonal anti-acetylated-α-tubulin (Sigma-Aldrich, St Louis, MO, USA); rabbit polyclonal anti-human Dync1h1 (amino-terminal 321 amino acids; serum #46, gift from Dr Richard Vallee, Columbia University); rabbit polyclonal anti-human Dync1h1 (carboxy-terminal 400 amino acids; 12345-1, ProteinTech, Chicago, IL, USA); rabbit polyclonal anti-rat Dync1h1 (amino acids 4,320 to 4,644; Dynein HC, R-325: sc-9115, Santa Cruz Biotechnology, Santa Cruz, CA, USA); rabbit polyclonal anti-Dync1i1 (gift from Dr Richard Vallee, Columbia University); rabbit polyclonal anti-Dynct1/p150 antibody (gift from Dr Kevin Vaughan, University of Notre Dame); mouse monoclonal anti-SV2 (Developmental Studies Hybridoma Bank).

Techniques: Control, Mutagenesis

Journal: Neural Development

Article Title: Analysis of a zebrafish dync1h1 mutant reveals multiple functions for cytoplasmic dynein 1 during retinal photoreceptor development

doi: 10.1186/1749-8104-5-12

Figure Lengend Snippet: Immunolocalization of Dync1h1 in developing photoreceptors . (A-E) Cryosections of the photoreceptor layer from wild-type retina (A, C, E) or cnb mutant retina (B, D) each at 4 dpf. Antisera directed against two different regions of Dync1h1 was used: (A, B) anti-human DYNC1H1 (huDYNC1H1; carboxy-terminal 400 amino acids) and (C, D) anti-rat Dync1h1 (amino acids 4,320 to 4,644). Primary antibodies were omitted in (E). For each panel, the fluorescent image is shown on the left and the transmitted light image on the right. CB, cell body; OS, outer segment; RPE, retina pigment epithelium; SR, synaptic region.

Article Snippet: Antibodies included: mouse monoclonal anti-acetylated-α-tubulin (Sigma-Aldrich, St Louis, MO, USA); rabbit polyclonal anti-human Dync1h1 (amino-terminal 321 amino acids; serum #46, gift from Dr Richard Vallee, Columbia University); rabbit polyclonal anti-human Dync1h1 (carboxy-terminal 400 amino acids; 12345-1, ProteinTech, Chicago, IL, USA); rabbit polyclonal anti-rat Dync1h1 (amino acids 4,320 to 4,644; Dynein HC, R-325: sc-9115, Santa Cruz Biotechnology, Santa Cruz, CA, USA); rabbit polyclonal anti-Dync1i1 (gift from Dr Richard Vallee, Columbia University); rabbit polyclonal anti-Dynct1/p150 antibody (gift from Dr Kevin Vaughan, University of Notre Dame); mouse monoclonal anti-SV2 (Developmental Studies Hybridoma Bank).

Techniques: Mutagenesis

Journal: Neural Development

Article Title: Analysis of a zebrafish dync1h1 mutant reveals multiple functions for cytoplasmic dynein 1 during retinal photoreceptor development

doi: 10.1186/1749-8104-5-12

Figure Lengend Snippet: Dync1h1, Dync1i1, and Dnct1 immunoreactivity is associated with photoreceptor axonemes . Immunocytochemical localization of proteins in isolated adult zebrafish photoreceptor cells. (A) Immunocytochemical localization of Dynein1 heavy chain (Dync1h1) using antisera directed against three different epitopes. (B) Immunocytochemical localization of Dynein1 intermediate chain (Dync1i1), (C) Dynactin1/p150 (Dctn1), and (D) Synaptic vesicle protein 2 (SV2). (E) Immunostaining in which the primary antibody was omitted. In (A-C), cells were co-stained with acetylated α-tubulin (Tub) to highlight the axoneme. Color merged (Merge) and transmitted light images overlaid with Merge images (Trans) are shown for each panel. IS, inner segment; OS, outer segment. NP, no primary antibody. (F) Western blot analysis on detergent-extracted photoreceptor cytoskeleton (DEPC) fraction shows the specificity of the Dync1h1 antibody, as the Dync2 h2 antibody recognizes a discrete and faster migrating band.

Article Snippet: Antibodies included: mouse monoclonal anti-acetylated-α-tubulin (Sigma-Aldrich, St Louis, MO, USA); rabbit polyclonal anti-human Dync1h1 (amino-terminal 321 amino acids; serum #46, gift from Dr Richard Vallee, Columbia University); rabbit polyclonal anti-human Dync1h1 (carboxy-terminal 400 amino acids; 12345-1, ProteinTech, Chicago, IL, USA); rabbit polyclonal anti-rat Dync1h1 (amino acids 4,320 to 4,644; Dynein HC, R-325: sc-9115, Santa Cruz Biotechnology, Santa Cruz, CA, USA); rabbit polyclonal anti-Dync1i1 (gift from Dr Richard Vallee, Columbia University); rabbit polyclonal anti-Dynct1/p150 antibody (gift from Dr Kevin Vaughan, University of Notre Dame); mouse monoclonal anti-SV2 (Developmental Studies Hybridoma Bank).

Techniques: Isolation, Immunostaining, Staining, Western Blot

Journal: Neural Development

Article Title: Analysis of a zebrafish dync1h1 mutant reveals multiple functions for cytoplasmic dynein 1 during retinal photoreceptor development

doi: 10.1186/1749-8104-5-12

Figure Lengend Snippet: Knock-down of Dync1h1 phenocopies cnb mutants . (A-C) TEM of retinas from 3-dpf embryos injected with a high dose (8 ng/embryo) of dync1h1 morpholino (MO). Photoreceptors across the entire retina failed to form outer segments and had rounded nuclei (N). (D-L) TEM views of embryos injected with a lower amount (3 ng/embryo) of dyn1 hc1 morpholino. Outer segments formed in some photoreceptors at the central (D, E) and peripheral retina (F) (arrows). Most lower dose morphant cells still failed to elongate outer segments. Scale bars: 10 μm in (A-E); 2 μm in (F).

Article Snippet: Antibodies included: mouse monoclonal anti-acetylated-α-tubulin (Sigma-Aldrich, St Louis, MO, USA); rabbit polyclonal anti-human Dync1h1 (amino-terminal 321 amino acids; serum #46, gift from Dr Richard Vallee, Columbia University); rabbit polyclonal anti-human Dync1h1 (carboxy-terminal 400 amino acids; 12345-1, ProteinTech, Chicago, IL, USA); rabbit polyclonal anti-rat Dync1h1 (amino acids 4,320 to 4,644; Dynein HC, R-325: sc-9115, Santa Cruz Biotechnology, Santa Cruz, CA, USA); rabbit polyclonal anti-Dync1i1 (gift from Dr Richard Vallee, Columbia University); rabbit polyclonal anti-Dynct1/p150 antibody (gift from Dr Kevin Vaughan, University of Notre Dame); mouse monoclonal anti-SV2 (Developmental Studies Hybridoma Bank).

Techniques: Knockdown, Injection

Journal: Neural Development

Article Title: Analysis of a zebrafish dync1h1 mutant reveals multiple functions for cytoplasmic dynein 1 during retinal photoreceptor development

doi: 10.1186/1749-8104-5-12

Figure Lengend Snippet: Summary of photoreceptor phenotypes associated with loss of Dynein1 or Dynactin

Article Snippet: Antibodies included: mouse monoclonal anti-acetylated-α-tubulin (Sigma-Aldrich, St Louis, MO, USA); rabbit polyclonal anti-human Dync1h1 (amino-terminal 321 amino acids; serum #46, gift from Dr Richard Vallee, Columbia University); rabbit polyclonal anti-human Dync1h1 (carboxy-terminal 400 amino acids; 12345-1, ProteinTech, Chicago, IL, USA); rabbit polyclonal anti-rat Dync1h1 (amino acids 4,320 to 4,644; Dynein HC, R-325: sc-9115, Santa Cruz Biotechnology, Santa Cruz, CA, USA); rabbit polyclonal anti-Dync1i1 (gift from Dr Richard Vallee, Columbia University); rabbit polyclonal anti-Dynct1/p150 antibody (gift from Dr Kevin Vaughan, University of Notre Dame); mouse monoclonal anti-SV2 (Developmental Studies Hybridoma Bank).

Techniques:

Journal: Neural Development

Article Title: Analysis of a zebrafish dync1h1 mutant reveals multiple functions for cytoplasmic dynein 1 during retinal photoreceptor development

doi: 10.1186/1749-8104-5-12

Figure Lengend Snippet: Dync1h1 knock-down causes dose-sensitive outer segment defects . TEM of retinas from 3-dpf embryos injected with a high dose (8 ng/embryo) of dync1h1 morpholino (MO). (A) A cone cell with Golgi apparatus (G) localized in the central ellipsoid region and mitochondria (M) located near the nucleus (N). (B-I) Cone cells with accumulation of vesicles (arrows) at the base of the connecting cilium (CC) and presence of disorganized disc structures within the outer segment (OS; arrowheads). (D-F) Severely disrupted outer segments showing vesiculation and tubulation of discs structures distally (arrowheads), yet normal discs at the base. Large vacuoles and vesicle accumulations were found at the base of the connecting cilium (arrows). The condensed cell in (F) is consistent with the morphology of early stage apoptosis. (G-I) Higher magnification insets of the red boxed regions. Scale bars: 0.5 μm in (A-F).

Article Snippet: Antibodies included: mouse monoclonal anti-acetylated-α-tubulin (Sigma-Aldrich, St Louis, MO, USA); rabbit polyclonal anti-human Dync1h1 (amino-terminal 321 amino acids; serum #46, gift from Dr Richard Vallee, Columbia University); rabbit polyclonal anti-human Dync1h1 (carboxy-terminal 400 amino acids; 12345-1, ProteinTech, Chicago, IL, USA); rabbit polyclonal anti-rat Dync1h1 (amino acids 4,320 to 4,644; Dynein HC, R-325: sc-9115, Santa Cruz Biotechnology, Santa Cruz, CA, USA); rabbit polyclonal anti-Dync1i1 (gift from Dr Richard Vallee, Columbia University); rabbit polyclonal anti-Dynct1/p150 antibody (gift from Dr Kevin Vaughan, University of Notre Dame); mouse monoclonal anti-SV2 (Developmental Studies Hybridoma Bank).

Techniques: Knockdown, Injection

Journal: Neural Development

Article Title: Analysis of a zebrafish dync1h1 mutant reveals multiple functions for cytoplasmic dynein 1 during retinal photoreceptor development

doi: 10.1186/1749-8104-5-12

Figure Lengend Snippet: Localization of Golgi apparatus in dync1h1 morphants . (A-F) Expression of a tol2-based gnat2: Man2a(1-100aa)-GFP construct in either control morphants (3 ng/embryo) (A-C) or dync1h1 morphants (3 ng/embryo) (D-F). Cryosections of the photoreceptor (PhR) layer at 4 dpf is shown. Embryos were treated with phenylthiourea to block pigment synthesis. Note that disorganization and mis-positioning of the Golgi apparatus in dync1h1 morphants. CB, cell body; IS, inner segment; MO, morpholino; OS, outer segment; RPE, retina pigment epithelium; SR, synaptic region.

Article Snippet: Antibodies included: mouse monoclonal anti-acetylated-α-tubulin (Sigma-Aldrich, St Louis, MO, USA); rabbit polyclonal anti-human Dync1h1 (amino-terminal 321 amino acids; serum #46, gift from Dr Richard Vallee, Columbia University); rabbit polyclonal anti-human Dync1h1 (carboxy-terminal 400 amino acids; 12345-1, ProteinTech, Chicago, IL, USA); rabbit polyclonal anti-rat Dync1h1 (amino acids 4,320 to 4,644; Dynein HC, R-325: sc-9115, Santa Cruz Biotechnology, Santa Cruz, CA, USA); rabbit polyclonal anti-Dync1i1 (gift from Dr Richard Vallee, Columbia University); rabbit polyclonal anti-Dynct1/p150 antibody (gift from Dr Kevin Vaughan, University of Notre Dame); mouse monoclonal anti-SV2 (Developmental Studies Hybridoma Bank).

Techniques: Expressing, Construct, Control, Blocking Assay

Journal: Neural Development

Article Title: Analysis of a zebrafish dync1h1 mutant reveals multiple functions for cytoplasmic dynein 1 during retinal photoreceptor development

doi: 10.1186/1749-8104-5-12

Figure Lengend Snippet: Ultrastructure of synaptic region in cnb mutants and dync1h1 morphants . TEMs of the photoreceptor synaptic region and outer plexiform layer of 3-dpf embryos. (A) Control morphants (3 ng/embryo) showed well developed synaptic input invaginations (asterisks) and numerous floating ribbons (arrows). (B) dync1h1 morphants (3 ng/embryo) had less defined synaptic input invaginations (asterisk) and significantly reduced number of floating ribbons (arrow). (C) cnb mutants did not show synaptic input invaginations and typically lacked ribbons. MO, morpholino.

Article Snippet: Antibodies included: mouse monoclonal anti-acetylated-α-tubulin (Sigma-Aldrich, St Louis, MO, USA); rabbit polyclonal anti-human Dync1h1 (amino-terminal 321 amino acids; serum #46, gift from Dr Richard Vallee, Columbia University); rabbit polyclonal anti-human Dync1h1 (carboxy-terminal 400 amino acids; 12345-1, ProteinTech, Chicago, IL, USA); rabbit polyclonal anti-rat Dync1h1 (amino acids 4,320 to 4,644; Dynein HC, R-325: sc-9115, Santa Cruz Biotechnology, Santa Cruz, CA, USA); rabbit polyclonal anti-Dync1i1 (gift from Dr Richard Vallee, Columbia University); rabbit polyclonal anti-Dynct1/p150 antibody (gift from Dr Kevin Vaughan, University of Notre Dame); mouse monoclonal anti-SV2 (Developmental Studies Hybridoma Bank).

Techniques: Control

Journal: Neural Development

Article Title: Analysis of a zebrafish dync1h1 mutant reveals multiple functions for cytoplasmic dynein 1 during retinal photoreceptor development

doi: 10.1186/1749-8104-5-12

Figure Lengend Snippet: Efficient transport of Rhodopsin(CT44)-GFP to the outer segment is disrupted with loss of Dync1h1 or Dctn1 . Cryosections of 3.5-dpf retina from transgenic Tg(1.3 xops :xRhoCT44-GFP) embryos in which the carboxy-terminal 44 amino acids of Rhodopsin are fused with GFP. Localization of GFP was evaluated in photoreceptors from (A) control morphants, (B) cnb mutants, (C) dync1h1 morphants or (D) dctn1a / b morphants. Upper row shows propidium iodide staining to highlight nuclei. Middle row shows RhoCT44-GFP. Bottom row shows color combined images. Arrowheads indicate cells with mislocalized GFP to the cell body region.

Article Snippet: Antibodies included: mouse monoclonal anti-acetylated-α-tubulin (Sigma-Aldrich, St Louis, MO, USA); rabbit polyclonal anti-human Dync1h1 (amino-terminal 321 amino acids; serum #46, gift from Dr Richard Vallee, Columbia University); rabbit polyclonal anti-human Dync1h1 (carboxy-terminal 400 amino acids; 12345-1, ProteinTech, Chicago, IL, USA); rabbit polyclonal anti-rat Dync1h1 (amino acids 4,320 to 4,644; Dynein HC, R-325: sc-9115, Santa Cruz Biotechnology, Santa Cruz, CA, USA); rabbit polyclonal anti-Dync1i1 (gift from Dr Richard Vallee, Columbia University); rabbit polyclonal anti-Dynct1/p150 antibody (gift from Dr Kevin Vaughan, University of Notre Dame); mouse monoclonal anti-SV2 (Developmental Studies Hybridoma Bank).

Techniques: Transgenic Assay, Control, Staining

Journal: Neural Development

Article Title: Analysis of a zebrafish dync1h1 mutant reveals multiple functions for cytoplasmic dynein 1 during retinal photoreceptor development

doi: 10.1186/1749-8104-5-12

Figure Lengend Snippet: Schematic of dose-dependent defects observed with loss of Dync1h1 . Illustrations of 3-dpf zebrfish cone photoreceptors from wild-type, low-dose dync1h1 morphants, and cnb mutants. Wild-type photoreceptors typically showed non-vesiculated outer segments, well organized connecting cilium and basal body, tightly packed mitochondria in the ellipsoid region, Golgi apparatus just apical to the nucleus in the myoid region, and numerous inputs and floating ribbons in the synaptic region. Photoreceptors of dync1h1 morphants and cnb mutants showed a spectrum of phenotypes with the most normal photoreceptors positioned in the central retina and more severely affected photoreceptors at the periphery. Cellular defects included increased incidence of outer segment vesiculation, varying degrees of organelle dispersion and depolarization, and under-development of the synaptic region. OS, outer segment; IS, inner segment; CB cell body; SR, synaptic region; E, ellipsoid region; M, myoid region.

Article Snippet: Antibodies included: mouse monoclonal anti-acetylated-α-tubulin (Sigma-Aldrich, St Louis, MO, USA); rabbit polyclonal anti-human Dync1h1 (amino-terminal 321 amino acids; serum #46, gift from Dr Richard Vallee, Columbia University); rabbit polyclonal anti-human Dync1h1 (carboxy-terminal 400 amino acids; 12345-1, ProteinTech, Chicago, IL, USA); rabbit polyclonal anti-rat Dync1h1 (amino acids 4,320 to 4,644; Dynein HC, R-325: sc-9115, Santa Cruz Biotechnology, Santa Cruz, CA, USA); rabbit polyclonal anti-Dync1i1 (gift from Dr Richard Vallee, Columbia University); rabbit polyclonal anti-Dynct1/p150 antibody (gift from Dr Kevin Vaughan, University of Notre Dame); mouse monoclonal anti-SV2 (Developmental Studies Hybridoma Bank).

Techniques: Dispersion