Journal: Molecular neurobiology

Article Title: Microtubule-Actin Crosslinking Factor 1 is required for dendritic arborization and axon outgrowth in the developing brain

doi: 10.1007/s12035-015-9508-4

Figure Lengend Snippet: (A) MACF1 expression in cortical neurons. Cortical neurons from E14.5 control (MACF1loxP/+; Nex-cre) and MACF1loxP/loxP; Nex-cre brains were cultured for 5 days, and MACF1 localization was examined using immunostaining. Arrows indicate an accumulation of MACF1 in dendritic tips (B) MACF1 deletion causes aberrant actin arrangement and localization. Polymerized actins were visualized by phalloidin staining. Top panels show the patterns of polymerized actins at neurite tips. Middel and bottom panels show polymerized actins accumulated in the cytosol. (C) The levels of polymerized actin (F-actin) intensity in the cell body and at the neurite tip were quantified using Image J (NIH) software. n = 21 cells from 3 independent cultures using 3 mice for each condition. Statistical significance was determined by two-tailed Student's t-test. ***p < 0.001. (D) MACF1-deleted neurons show abnormal microtubule arrangement at the neurite tip. (E) Immunoblotting was performed to measure the levels of α-tubulin or acetylated-tubulin using E14.5 control and MACF1loxP/loxP; Nex-cre brain lysates (top panel). The levels of each tubulin were quantified in the bottom panel. n = 3 blots using 3 different lysates from 3 mice for each condition. Statistical significance was determined by two-tailed Student's t-test. ***p < 0.001.

Article Snippet: The following primary antibodies were used: Chicken anti-GFP (Invitrogen), Rabbit anti-GFP (Invitrogen), mouse anti-β-III Tubulin (Phosphosolutions), rabbit anti-acetyl-α-tubulin (Cell Signaling), and mouse anti-β-tubulin (Upstate).

Techniques: Expressing, Control, Cell Culture, Immunostaining, Staining, Software, Two Tailed Test, Western Blot

Journal: Nanoscale Research Letters

Article Title: A nerve guidance conduit with topographical and biochemical cues: potential application using human neural stem cells

doi: 10.1186/s11671-015-0972-6

Figure Lengend Snippet: Fluorescence microscopy images of hNSCs cultured with PCL conduit for 14 days: a DAPI and b β-III tubulin. DAPI staining is indicated in blue and β-III tubulin staining is indicated in green. Arrow indicates the direction of aligned nanofibers

Article Snippet: After cryosectioning (20-μm slices), each section was washed twice in PBS for 10 min. Then, the samples were treated with blocking buffer (3 % goat serum PBS with 0.01 % Triton X-100) for 1 h at room temperature before adding β-III tubulin goat anti-mouse primary antibody (1:200 dilution, Promega, Madison, WI, USA) overnight at 4 °C.

Techniques: Fluorescence, Microscopy, Cell Culture, Staining

Journal: Nanoscale Research Letters

Article Title: A nerve guidance conduit with topographical and biochemical cues: potential application using human neural stem cells

doi: 10.1186/s11671-015-0972-6

Figure Lengend Snippet: Confocal microscopy images of hNSCs cultured with PSHU-RGD/PCL conduit for 14 days: representative images a , b along the center and e , f inner wall of microchannels, a , d DAPI, b , e β-III tubulin, and c , f combined DAPI and β-III tubulin images. DAPI staining is indicated in blue and β-III tubulin staining is indicated in green . Images show that hNSCs migrated far into the conduit with extensive neurite extension along the nanofiber direction ( arrows )

Article Snippet: After cryosectioning (20-μm slices), each section was washed twice in PBS for 10 min. Then, the samples were treated with blocking buffer (3 % goat serum PBS with 0.01 % Triton X-100) for 1 h at room temperature before adding β-III tubulin goat anti-mouse primary antibody (1:200 dilution, Promega, Madison, WI, USA) overnight at 4 °C.

Techniques: Confocal Microscopy, Cell Culture, Staining

Journal: Frontiers in Immunology

Article Title: Antibody Repertoires Identify β-Tubulin as a Host Protective Parasite Antigen in Mice Infected With Trypanosoma cruzi

doi: 10.3389/fimmu.2018.00671

Figure Lengend Snippet: Infection induces a distinct antibody repertoire: a major reactivity elicited against Trypanosoma cruzi is directed to T. cruzi β-tubulin. (A) Immunoblot showing individual serum IgG reactivities from control ( n = 3) and chronically infected BALB/c mice ( n = 4) against T. cruzi extracts. The left arrow indicates the major 50–55 kDa protein band in the protein extract. IgG concentrations of all sera were adjusted to 100 µg/mL. (B) Left lane: Coomassie blue staining of SDS-PAGE showing the protein profile of T. cruzi extract. (B) Right lane: immunoblot of recombinant TcβTUB probed with anti-mouse β-tubulin III monoclonal antibody 5G8.

Article Snippet: Immunoblots against recombinant TcβTUB were performed with either anti-mouse β-tubulin III monoclonal antibody 5G8 (Promega) or anti-β-tubulin rabbit monoclonal antibody 9F3 (Cell Signaling).

Techniques: Infection, Western Blot, Staining, SDS Page, Recombinant

Journal: Frontiers in Immunology

Article Title: Antibody Repertoires Identify β-Tubulin as a Host Protective Parasite Antigen in Mice Infected With Trypanosoma cruzi

doi: 10.3389/fimmu.2018.00671

Figure Lengend Snippet: Global serum IgG reactivities against Trypanosoma cruzi and autologous heart polypeptides in infected BALB/c mice with T. cruzi . Mean densitometric profiles of IgG reactivities during acute (A) or chronic (B) infection on immunoblots against T. cruzi (left) and autologous heart polypeptides (right). Individual serum IgG concentrations were adjusted to 100 µg/mL. Arrow indicates the most prominent band of reactivity observed in the sera of acutely and chronically infected mice. Densitometric profiles represent the mean of 5–8 individual sera (see ) (** p < 0.01; *** p < 0.001). Immunoblots against heart polypeptides did not show significant differences. Data are representative of at least three independent experiments with five to eight mice per group. (C) A representative immunoblot of chronic serum against T. cruzi extract. Arrows indicate the three major T. cruzi protein bands, which were identified by mass spectrometry analysis as pyruvate phosphate dikinase (100 kDa), Hsp-85 (85 kDa), and β-tubulin (55 kDa). We could not identify the small-molecular weight band that also displayed a pronounced peak.

Article Snippet: Immunoblots against recombinant TcβTUB were performed with either anti-mouse β-tubulin III monoclonal antibody 5G8 (Promega) or anti-β-tubulin rabbit monoclonal antibody 9F3 (Cell Signaling).

Techniques: Infection, Western Blot, Mass Spectrometry, Molecular Weight

Journal: Frontiers in Immunology

Article Title: Antibody Repertoires Identify β-Tubulin as a Host Protective Parasite Antigen in Mice Infected With Trypanosoma cruzi

doi: 10.3389/fimmu.2018.00671

Figure Lengend Snippet: Mass spectrometry analysis of peptide fragments of the major 50–55 kDa band of Trypanosoma cruzi extract corresponding to T. cruzi β-tubulin.

Article Snippet: Immunoblots against recombinant TcβTUB were performed with either anti-mouse β-tubulin III monoclonal antibody 5G8 (Promega) or anti-β-tubulin rabbit monoclonal antibody 9F3 (Cell Signaling).

Techniques: Mass Spectrometry, Sequencing

Journal: Frontiers in Immunology

Article Title: Antibody Repertoires Identify β-Tubulin as a Host Protective Parasite Antigen in Mice Infected With Trypanosoma cruzi

doi: 10.3389/fimmu.2018.00671

Figure Lengend Snippet: Infection with Trypanosoma cruzi increases IgG reactivity against T. cruzi β-tubulin. Serum IgG reactivities from control uninfected (open circles) or chronically infected (closed circles) BALB/c mice were assayed by ELISA on plates coated with recombinant TcβTUB. All IgG concentrations in sera were adjusted to 100 µg/mL. Each symbol corresponds to one individual mouse. Statistical analysis was performed by t -test from representative results of three similar experiments (** p < 0.01).

Article Snippet: Immunoblots against recombinant TcβTUB were performed with either anti-mouse β-tubulin III monoclonal antibody 5G8 (Promega) or anti-β-tubulin rabbit monoclonal antibody 9F3 (Cell Signaling).

Techniques: Infection, Enzyme-linked Immunosorbent Assay, Recombinant

Journal: Frontiers in Immunology

Article Title: Antibody Repertoires Identify β-Tubulin as a Host Protective Parasite Antigen in Mice Infected With Trypanosoma cruzi

doi: 10.3389/fimmu.2018.00671

Figure Lengend Snippet: Increased IgG reactivity against β-tubulin in mice immunized with recombinant TcβTUB. (A) Immunoblots from mice immunized with BSA/CFA before infection and (B) mice immunized with TcβTUB/CFA before infection. Sera were blotted against mouse tubulin from mouse brain extract (MBE), recombinant TcβTUB and native tubulin from Trypanosoma cruzi extract. (C) Immunoblots of MBE, recombinant TcβTUB, and T. cruzi extract with monoclonal antibodies against α-tubulin; and (D) against β-tubulin. The shift in molecular weight of recombinant TcβTUB, compared with native mouse and T. cruzi β-tubulins, is due to an additional poly-histidine tail. (E) Densitometric profiles of the staining profiles presented in panels (A,B) .

Article Snippet: Immunoblots against recombinant TcβTUB were performed with either anti-mouse β-tubulin III monoclonal antibody 5G8 (Promega) or anti-β-tubulin rabbit monoclonal antibody 9F3 (Cell Signaling).

Techniques: Recombinant, Western Blot, Infection, Molecular Weight, Staining

Journal: Anatomical record (Hoboken, N.J. : 2007)

Article Title: Comparative morphology and histology of the nasal fossa in four mammals: gray squirrel, bobcat, coyote and white-tailed deer

doi: 10.1002/ar.23352

Figure Lengend Snippet: Epithelial lining of the bobcat nasal fossa. Sagittal view of segmented MRI scans of the bobcat nasal airway showing five coronal sections (b–f) selected for morphological and histological representation (A). The five coronal sections along the rostrocaudal axis illustrate the maxilloturbinal (B–C), nasomaxillary (D) and ethmoturbinal (E–F) regions of the bobcat nasal fossa. Magnified views show the nasoturbinal (G), maxilloturbinal (H, J) and ethmoturbinal (I) covered with nonsensory epithelium and packed with Alcian blue labeled goblet cells at rostral regions of the nasal fossa. Olfactory epithelium covered the nasoturbinal (K), septum (L), and ethmoturbinal (L) in more caudal regions and had the characteristic epithelial thickness and Bowman’s glands in the lamina propria (M) and labeled with β-tubulin III antibody (N–O). nt = nasoturbinal; mt = maxilloturbinal; s = septum; et = ethmoturbinals; ob = olfactory bulb; oe = olfactory epithelium; bg = Bowman’s gland; ne = nonsensory epithelium. Scale bar: B–F = 2 mm; G–L, N = 100 µm; and M, O = 50 µm.

Article Snippet: To further visualize the OE, we conducted immunohistochemistry with a mouse monoclonal anti-neuronal class III β-tubulin antibody (TMJ1, Babco) as has been used to label olfactory neurons in rodents ( Roskams et al., 1998 ), cats (Lishcka et al., 2008) and humans ( Ronnett et al., 2003 ), suggesting reactivity of this antibody in diverse mammalian samples.

Techniques: Labeling

Journal: Anatomical record (Hoboken, N.J. : 2007)

Article Title: Comparative morphology and histology of the nasal fossa in four mammals: gray squirrel, bobcat, coyote and white-tailed deer

doi: 10.1002/ar.23352

Figure Lengend Snippet: Epithelial lining of the coyote nasal fossa. Sagittal view of segmented MRI scans of the coyote nasal airway showing five coronal sections (b–f) selected for morphological and histological representation (A). The five coronal sections along the rostrocaudal axis illustrate the maxilloturbinal (B–C), nasomaxillary (D) and ethmoturbinals (E–F) regions of the coyote nasal fossa. Nonsensory epithelium covered the nasoturbinal (G), maxilloturbinal (H–K), and ethmoturbinal (L) at the rostral regions. Thick olfactory epithelium (L), clearly labeled with β-tubulin antibody (N–O), covered the septum, nasoturbinal, and ethmoturbinal at the caudal regions of the nasal fossa. nt = nasoturbinal; mt = maxilloturbinal; s = septum; et = ethmoturbinal; fs = frontal sinus; ob = olfactory bulb; oe = olfactory epithelium; ne = nonsensory epithelium. Scale bar: B–F = 2 mm; G–L, N = 100 µm; and M, O = 50 µm.

Article Snippet: To further visualize the OE, we conducted immunohistochemistry with a mouse monoclonal anti-neuronal class III β-tubulin antibody (TMJ1, Babco) as has been used to label olfactory neurons in rodents ( Roskams et al., 1998 ), cats (Lishcka et al., 2008) and humans ( Ronnett et al., 2003 ), suggesting reactivity of this antibody in diverse mammalian samples.

Techniques: Labeling