anti β tubulin iii tuj1  (Neuromics)

Journal: ACS Omega

Article Title: LSD Modulates Proteins Involved in Cell Proteostasis, Energy Metabolism and Neuroplasticity in Human Cerebral Organoids

doi: 10.1021/acsomega.4c04712

Figure Lengend Snippet: Characterization of the human cerebral organoids. Nuclei are stained blue with 4′,6-Diamidino-2-Phenylindole, Dihydrochloride (DAPI) in all images. (A) Immunostaining for 5-HT 2A receptors, which colocalize with the neuronal marker MAP2; (B) TUJ1 immunostaining (green), a neuronal marker, accompanied by PAX6 immunostaining in red, which highlights neural progenitors; (C) GFAP immunostaining identifies radial glia and astrocytes. Scale bars represent 400 μm for whole organoid images and 60 μm for zoomed-in images.

Article Snippet: Subsequently, a blocking step was carried out by incubating in a solution containing 3% fetal calf serum in PBS for 1 h. Following blocking, they were subjected to overnight incubation at 4°C with the primary antibody TUJ1 (Neuromics) at a dilution of 1:2000 in the blocking solution.

Techniques: Staining, Immunostaining, Marker

Journal: ACS Omega

Article Title: LSD Modulates Proteins Involved in Cell Proteostasis, Energy Metabolism and Neuroplasticity in Human Cerebral Organoids

doi: 10.1021/acsomega.4c04712

Figure Lengend Snippet: Neurite Outgrowth in Response to LSD: Human brain spheroids were initially plated for 24 h and then subjected to exposure to either 10 or 100 nM LSD for an additional 24 h. For this analysis, the control group included 18 spheroids, while the groups exposed to 10 and 100 nM LSD consisted of 17 and 15 spheroids, respectively. (A) The samples were analyzed using immunofluorescence staining for β-tubulin III (TUJ1; depicted in green) and DAPI staining to highlight nuclei (shown in blue). (B) Sholl analysis was employed to assess neurite outgrowth. An inset in this section provides a schematic of the Sholl analysis method. The analysis was conducted on the mean numbers of crossings for each group of five circles. The quantification was carried out over five independent experiments. Statistical significance is indicated as follows: * p < 0.05, ** p < 0.01, *** p < 0.001, in comparison to the control group. A scale bar indicating 1000 μm is included for reference.

Article Snippet: Subsequently, a blocking step was carried out by incubating in a solution containing 3% fetal calf serum in PBS for 1 h. Following blocking, they were subjected to overnight incubation at 4°C with the primary antibody TUJ1 (Neuromics) at a dilution of 1:2000 in the blocking solution.

Techniques: Control, Immunofluorescence, Staining, Comparison

Journal: JCI Insight

Article Title: Gut mucosal cells transfer α -synuclein to the vagus nerve

doi: 10.1172/jci.insight.172192

Figure Lengend Snippet: ( A ) Intestinal organoids were prepared from an SNCA A53T mouse in which CCK-containing cells express enhanced green fluorescent protein (eGFP), and vagal nodose ganglia neurons were isolated from an Snca –/– mouse lacking endogenous α-synuclein. ( B ) Representative images of organoids and neurons grown in coculture for 5 days, with eGFP-positive cells (green) in the organoid and β-tubulin III (Tuj1, cyan) highlighting neuronal processes. ( C ) Representative high-magnification α-synuclein (red) staining of an eGFP-positive EEC. Red arrow indicates localization to a PGP9.5-positive (cyan) process in an Snca –/– mouse neuron. ( D and E ) Representative images with neuron-specific β-tubulin III (cyan). Surface and (adjacent) intracellular confocal slices are shown. Scale bars are 30 μm for B , 3 μm for C , and 5 μm for D and E .

Article Snippet: Primary antibodies used for immunostaining included rabbit CCK , rabbit α-synuclein (Abcam catalog ab138501, RRID:AB_2537217, at 1:1,000), guinea pig PGP9.5 (Abcam catalog ab10410, RRID:AB_297150, at 1:100), chick β-tubulin III (Tuj1; Neuromics catalog CH23005, RRID:AB_2210684, at 1:100), and chick GFP (Abcam catalog ab13970, RRID:AB_300798, at 1:1,000).

Techniques: Isolation, Staining

Journal: Developmental biology

Article Title: Galnt17 loss-of-function leads to developmental delay and abnormal coordination, activity, and social interactions with cerebellar vermis pathology

doi: 10.1016/j.ydbio.2022.08.002

Figure Lengend Snippet: (A) Tuj1 (left) and CalB (right) staining showed the morphology of Purkinje cells in WT (top) and Galnt17−/− (bottom) animals. Galnt17−/− mutants lost thick primary branches (arrowheads), and the primary branches were bifurcated (arrows) compared to WT mice at P60. (B) Galnt17−/− mutants (bottom) showed fewer CalB (green) marked axons in axonal tract in lobule IV/V compared to WT mice (top) at P14. (C) Width of axonal tract was measured, and it showed mutants (dark gray) have narrower axonal tract than WT (light gray) in lobule IV/V at P14. n=3 for each genotype. t-Test, *p<0.05. (D) Galnt17−/− mutants (right) showed forelimb clasping (arrowhead) during tail suspension test compared to WT (left) mice in adult. n=5 animals for each genotype were scored for clasping (1) or no clasping (0) with all mutant animals scoring as 1 and all WT animals scoring as 0 in this test. Scale bar = 50 μm.

Article Snippet: Primary antibodies: GFAP (Invitrogen 180063, 1:400), BrdU (Abcam ab6326, 1:400), Calbindin (CalB; Sigma C9848, 1:400), beta-III Tubulin (Tuj1; Neuromics CH23005, 1:200), Phospho-H3 (Millipore 06–570, 1:500); secondary antibodies: Thermo-Fisher Scientific, Alexa Fluor 488, Alexa Fluor 594.

Techniques: Staining, Suspension, Mutagenesis