Journal: Frontiers in Molecular Neuroscience

Article Title: Ptbp2 re-expression rescues axon growth defects in Smn-deficient motoneurons

doi: 10.3389/fnmol.2024.1393779

Figure Lengend Snippet: Ptbp2 interacts with Smn in motoneurons. (A) Representative images of Smn-Ptbp2 PLA signal in cultured motoneurons at DIV 6 using anti-Smn and anti-Ptbp2 antibodies. Motoneuron morphology was visualized with anti-Tubb3 antibody. Scale bars, 10 and 5 μm (magnified areas). (B) Co-immunoprecipitation of Smn by anti-Ptbp2 from motoneuron lysate pre-treated with RNase A as indicated. (C) Representative images showing Smn immunofluorescence and Hnrnpr FISH in cultured motoneurons at DIV 6. Arrowheads indicate colocalization of Smn and Hnrnpr in granules. Scale bars, 10 and 2 μm (magnified areas). (D) Fluorescence intensity profiles of Smn and Hnrnpr at the location indicated by arrow 4 in (C) . (E) Representative images showing Ptbp2 and Smn immunofluorescence and Hnrnpr FISH in cultured motoneurons at DIV 6. Scale bars, 10 and 2 μm (magnified areas). (F) Fluorescence intensity profiles of Ptbp2, Smn and Hnrnpr at the location indicated by a line in (E) .

Article Snippet: Cells were fixed again for 10 min at room temperature in PLP, washed with DPBS, and stained with FITC-conjugated anti-Tubb3 antibody (130-131-158, Miltenyi Biotec).

Techniques: Cell Culture, Immunoprecipitation, Immunofluorescence, Fluorescence

Journal: iScience

Article Title: Nuclear miR-150 enhances hepatic lipid accumulation by targeting RNA transcripts overlapping the PLIN2 promoter

doi: 10.1016/j.isci.2023.107837

Figure Lengend Snippet:

Article Snippet: Anti-Beta Tubulin Antibody , Boster , Cat# M01857-3.

Techniques: Recombinant, Magnetic Beads, SYBR Green Assay, Lysis, Luciferase, Reporter Assay, cDNA Synthesis, Plasmid Preparation, Software

Journal: Graefe's Archive for Clinical and Experimental Ophthalmology

Article Title: Neural stem/progenitor cells circulating in peripheral blood of patients with neovascular form of AMD: a novel view on pathophysiology

doi: 10.1007/s00417-011-1767-9

Figure Lengend Snippet: Identification of CXCR4 + Lin – CD45 - TCSCs derived from the peripheral blood of AMD patients. The individual images depict the expression of CXCR4 and β-III-tubulin ( a ), GFAP ( b ) and nestin ( c ) antigens in isolated CXCR4 + Lin – CD45 - cells. The nuclei were visualized via DAPI staining. A pseudocolour was assigned to each stain as follows: anti-CXCR4 – red , anti-nestin, β-III-tubulin and GFAP – green , nuclei – blue . The control images (the upper boxed insets ) confirm no unspecific binding of antibodies. The scale bar shows 5 μm. Representative data are shown

Article Snippet: The sorted cells were permeabilized (0.1% Triton X-100, 10 min) and stained for β-III-tubulin (OriGene Technologies, Rockville, MD, USA), nestin (Novus Biologicals, Littleton, CO, USA) and glial fibrillary acidic protein (GFAP) (Santa Cruz Biotechnology, Santa Cruz, CA, USA) antigens with anti-human monoclonal antibodies for 1 h at room temperature followed by incubation with a secondary antibody conjugated to FITC (Sigma, St. Louis, MO, USA).

Techniques: Derivative Assay, Expressing, Isolation, Staining, Binding Assay

Journal: Graefe's Archive for Clinical and Experimental Ophthalmology

Article Title: Neural stem/progenitor cells circulating in peripheral blood of patients with neovascular form of AMD: a novel view on pathophysiology

doi: 10.1007/s00417-011-1767-9

Figure Lengend Snippet: Expression of nestin, β-III-tubulin, and GFAP at the mRNA level in peripheral blood nuclear cells. Transcript quantity is expressed in relative units against the β2-microglobulin control gene (2 ΔCt , where Ct represents the threshold cycle difference between the control and target genes)

Article Snippet: The sorted cells were permeabilized (0.1% Triton X-100, 10 min) and stained for β-III-tubulin (OriGene Technologies, Rockville, MD, USA), nestin (Novus Biologicals, Littleton, CO, USA) and glial fibrillary acidic protein (GFAP) (Santa Cruz Biotechnology, Santa Cruz, CA, USA) antigens with anti-human monoclonal antibodies for 1 h at room temperature followed by incubation with a secondary antibody conjugated to FITC (Sigma, St. Louis, MO, USA).

Techniques: Expressing

Journal: Graefe's Archive for Clinical and Experimental Ophthalmology

Article Title: Neural stem/progenitor cells circulating in peripheral blood of patients with neovascular form of AMD: a novel view on pathophysiology

doi: 10.1007/s00417-011-1767-9

Figure Lengend Snippet: Correlation between the expression of some neuroprogenitor and glial cell markers in the study groups

Article Snippet: The sorted cells were permeabilized (0.1% Triton X-100, 10 min) and stained for β-III-tubulin (OriGene Technologies, Rockville, MD, USA), nestin (Novus Biologicals, Littleton, CO, USA) and glial fibrillary acidic protein (GFAP) (Santa Cruz Biotechnology, Santa Cruz, CA, USA) antigens with anti-human monoclonal antibodies for 1 h at room temperature followed by incubation with a secondary antibody conjugated to FITC (Sigma, St. Louis, MO, USA).

Techniques: Expressing

Journal: Investigative Ophthalmology & Visual Science

Article Title: Activation of Cytosolic Calpain, Not Caspase, Is Underlying Mechanism for Hypoxic RGC Damage in Human Retinal Explants

doi: 10.1167/iovs.61.13.13

Figure Lengend Snippet: Calpain activation in β-III tubulin-positive, NFL from human ( A ) and monkey ( B ) retinal explants cultured under hypoxia/reoxygenation. Images show confocal microscopy of flat mounts stained with SBDP150 ( red , left columns ) that were then merged with those stained for β-III tubulin ( green , right columns ). LABELS: Non-culture, before culture; N24 or N40, normoxia for 24 or 40 hours; H16R8, 16 hours hypoxia/8 hours reoxygenation; H24R16, 24 hours hypoxia/16 hours reoxygenation; +SNJ, cultured with 100 µM SNJ-1945 under each condition. Scale bar : 50 µm.

Article Snippet: Samples were stained with antibodies for SBDP150 as described above and β-III tubulin (as a neuron marker, 1:500 dilution; Merck).

Techniques: Activation Assay, Cell Culture, Confocal Microscopy, Staining

Journal: iScience

Article Title: C9orf72 poly-PR forms anisotropic condensates causative of nuclear TDP-43 pathology

doi: 10.1016/j.isci.2024.110937

Figure Lengend Snippet:

Article Snippet: Beta III Tubulin/Tuj-1 (chicken) , Merck , Cat# AB9354; RRID: AB_570918.

Techniques: Labeling, Virus, Recombinant, Transfection, Reverse Transcription, Lysis, Western Blot, Staining, Derivative Assay, Plasmid Preparation, Software, Imaging, Microscopy, Real-time Polymerase Chain Reaction

Journal: PLoS ONE

Article Title: No Dopamine Cell Loss or Changes in Cytoskeleton Function in Transgenic Mice Expressing Physiological Levels of Wild Type or G2019S Mutant LRRK2 and in Human Fibroblasts

doi: 10.1371/journal.pone.0118947

Figure Lengend Snippet: A-B: Neurite parameters of neuronal cultures from LRRK2, GS-LRRK2 (line 2) and their respective non-tg littermate, which were treated with vehicle-control or LRRK2-IN-1 (0.1 M) for seven days (DIV7). Comparison of parameters describing neurite branching (A) included number of branches, number of neurite trees and number of segments. Comparison of neurite length parameters (B) included total neurite length, average neurite length and maximal neurite length. Data represent mean ± SEM and were analyzed with two-way ANOVA. No significant difference was detected. Number of neurons analyzed for cultures obtained from LRRK2 transgenic mice: non-tg = 1339, non-tg + LRRK2-IN-1 = 1609; LRRK2 = 1697, LRRK2 + LRRK2-IN-1 = 1542, n = 4 independent experiments; Number of neurons analyzed for cultures obtained from GS-LRRK2 transgenic mice: non-tg = 1268; non-tg + LRRK2-IN-1 = 1522; GS-LRRK2 = 1526; GS-LRRK2 + LRRK2-IN-1 = 1844, n = 4 independent experiments; C-H: Representative pictures of ß-Tubulin III stained neurons on DIV7 derived from wild type, GS- LRRK2 (line 2), their non-transgenic littermates. Pictures were obtained with the BD Pathway 855 high content Bioimager. C1-H2: Total neurite length (C1-H1) and number of branches (C2-H2) segmentation corresponding to ß-tubulin III staining images (C-H) obtained from Attovision Software.

Article Snippet: At day in vitro (DIV) 3, 7 and 14 neuronal cultures were immunostained with mouse anti-ß-Tubulin, Class III antibody conjugated to Alexa Fluor 488 (1:50; BD Pharmingen) and the nuclear marker Hoechst 33342 (1:2000), both diluted in PBS.

Techniques: Transgenic Assay, Staining, Derivative Assay, Software

Journal: Pharmaceuticals

Article Title: The Role of TRPA1 Channels in the Central Processing of Odours Contributing to the Behavioural Responses of Mice

doi: 10.3390/ph14121336

Figure Lengend Snippet: Expression of Trpa1 mRNA in the investigated regions of the olfactory system of C57BL/6 mice ( n = 4). Trpa1 (red) mRNA signal did not colocalise with β-tubulin III (green) immunorective cells in the OE ( a ). Trpa1 (red) mRNA signal colocalised exclusively with NeuN (white) positive neurons in the OB ( b ). Trpa1 (red), Gad1 (white) and Vglut1 (green) mRNA expression in the OB (Bregma 3 mm), ( c ) and in the PC ( d ) (Bregma −1.46 mm). Trpa1 mRNA signal colocalised both with Gad1 and Vglut1 positive neurons in the OB ( c ), but it colocalised only with Vglut1 positive neurons in the PC ( d ). Cell nuclei were counterstained with DAPI (blue) in all areas. Abbreviations: NeuN: neuronal nuclear protein, Gad1: glutamate decarboxylase 1, Vglut1: vesicular glutamate transporter 1, DAPI: 4′,6-diamidino-2-phenylindole. In order to highlight the cell borders, differential interference contrast (DIC) images were merged with the virtual color images. Bars: 10 µm.

Article Snippet: Briefly, after channel development of the RNAscope assay, sections were washed for 2 × 15 min in PBS, incubated overnight at RT with polyclonal rabbit anti-β-tubulin III (Merck, Sigma Aldrich GmbH; Cat. No.: T2200, Schnelldorf, Germany), diluted 1:200 with 2% normal donkey serum for blocking aspecific binding.

Techniques: Expressing

Journal: Heliyon

Article Title: Cortical neurons obtained from patient-derived iPSCs with GNAO1 p.G203R variant show altered differentiation and functional properties

doi: 10.1016/j.heliyon.2024.e26656

Figure Lengend Snippet: Immunostaining analysis at day 25 of differentiation A-B . Immunostaining analysis with the indicated primary antibodies and DAPI to label nuclei. PAX6 and NESTIN are neural progenitor markers; TBR2 is a neuronal precursor marker; TUJ1 and MAP2 are neuronal markers.

Article Snippet: GNAO1 +/G203R differentiating cells were immunostained as described in Brighi et al., 2021 [ ] using the following primary antibodies: mouse anti-PAX6 (sc81649 Santa Cruz Biotechnology, 1:50), mouse anti-GFAP (MAB360 Merck Life Sciences, 1:500), chicken anti-MAP2 (ab5392 Abcam, 1:2000), rabbit anti-β-TUBULIN III (TUJ1) (T2200 Merck Life Sciences, 1:2000), Rabbit Anti-Nestin (MA5-32272 Thermo Fisher Scientific, 1:100), Rabbit Anti-TBR2 (ab15894 Millipore,1:50).

Techniques: Immunostaining, Marker

Journal: Heliyon

Article Title: Cortical neurons obtained from patient-derived iPSCs with GNAO1 p.G203R variant show altered differentiation and functional properties

doi: 10.1016/j.heliyon.2024.e26656

Figure Lengend Snippet: Analysis of neuronal rosettes morphology in the independent GNAO1 +/G203R #2 line A. Phase contrast images of GNAO1 +/G203R #2 cells at the neural rosette stage, corresponding to day 30 of differentiation. Note that the in the protocol used for differentiation of GNAO1 +/G203R #2 cells (as described in the Methods section) has a different timing from the one depicted in A. B-D. Immunostaining was performed using the specified primary antibodies along with DAPI for nuclear labeling. TBR1 is a marker of early-born neurons (B); and TUJ1 (B) and MAP2 (C) are markers for neurons; NESTIN serves as a marker for neural progenitors (C); β-catenin as a marker of apical polarization in neural rosettes (D). Number of individual cultures for experiments was 3.

Article Snippet: GNAO1 +/G203R differentiating cells were immunostained as described in Brighi et al., 2021 [ ] using the following primary antibodies: mouse anti-PAX6 (sc81649 Santa Cruz Biotechnology, 1:50), mouse anti-GFAP (MAB360 Merck Life Sciences, 1:500), chicken anti-MAP2 (ab5392 Abcam, 1:2000), rabbit anti-β-TUBULIN III (TUJ1) (T2200 Merck Life Sciences, 1:2000), Rabbit Anti-Nestin (MA5-32272 Thermo Fisher Scientific, 1:100), Rabbit Anti-TBR2 (ab15894 Millipore,1:50).

Techniques: Immunostaining, Labeling, Marker