Journal: BioMed Research International

Article Title: Bone Morphogenetic Protein-7 (BMP-7) Promotes Neuronal Differentiation of Bone Marrow Mesenchymal Stem Cells (BMSCs) In Vitro

doi: 10.1155/2021/7239783

Figure Lengend Snippet: Cell immunofluorescence detection. The expression and location of MAP-2 and Nfh were detected with the cell immunofluorescence (200x).

Article Snippet: After being treated with 0.5% Triton X-100 at room temperature for 20 min and the following washing, the cells were blocked with 5% BSA solution at room temperature for 20 min. Then, the cells were incubated with the rabbit anti-Nfh primary antibody (1 : 1500 dilution; bs-10680R; Bioss), or rabbit anti-MAP-2 primary antibody (1 : 1500 dilution; bs-1369R; Bioss), at 4°C overnight.

Techniques: Immunofluorescence, Expressing

Journal: BioMed Research International

Article Title: Bone Morphogenetic Protein-7 (BMP-7) Promotes Neuronal Differentiation of Bone Marrow Mesenchymal Stem Cells (BMSCs) In Vitro

doi: 10.1155/2021/7239783

Figure Lengend Snippet: Relative mRNA expression levels of MAP-2 and Nfh. (a, b) Quantitative real-time PCR was performed to detect the mRNA expression levels of MAP-2 (a) and Nfh (b). Compared with the control and lentiviral vector control groups, ∗ P < 0.05.

Article Snippet: After being treated with 0.5% Triton X-100 at room temperature for 20 min and the following washing, the cells were blocked with 5% BSA solution at room temperature for 20 min. Then, the cells were incubated with the rabbit anti-Nfh primary antibody (1 : 1500 dilution; bs-10680R; Bioss), or rabbit anti-MAP-2 primary antibody (1 : 1500 dilution; bs-1369R; Bioss), at 4°C overnight.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Plasmid Preparation

Journal: BioMed Research International

Article Title: Bone Morphogenetic Protein-7 (BMP-7) Promotes Neuronal Differentiation of Bone Marrow Mesenchymal Stem Cells (BMSCs) In Vitro

doi: 10.1155/2021/7239783

Figure Lengend Snippet: Relative protein expression levels of MAP-2 and Nfh. (a) Western blot analysis was performed to detect the protein expression levels of MAP-2 and Nfh. (b) Statistical analysis. Compared with the control and lentiviral vector control groups, ∗ P < 0.05.

Article Snippet: After being treated with 0.5% Triton X-100 at room temperature for 20 min and the following washing, the cells were blocked with 5% BSA solution at room temperature for 20 min. Then, the cells were incubated with the rabbit anti-Nfh primary antibody (1 : 1500 dilution; bs-10680R; Bioss), or rabbit anti-MAP-2 primary antibody (1 : 1500 dilution; bs-1369R; Bioss), at 4°C overnight.

Techniques: Expressing, Western Blot, Plasmid Preparation

Journal: iScience

Article Title: Dendritic spine formation and synapse maturation in transcription factor-induced human iPSC-derived neurons

doi: 10.1016/j.isci.2023.106285

Figure Lengend Snippet: Key resources table

Article Snippet: Rabbit polyclonal anti-MAP-2 , Millipore , Cat# AB5622-I; RRID: AB_2800501.

Techniques: Control, Recombinant, Membrane, Clone Assay, Reverse Transcription, RNA HS Assay, DNA Library Preparation, Expressing, Derivative Assay, Software, Microscopy, Cell Culture

Journal: PLoS ONE

Article Title: Murine Neural Stem/Progenitor Cells Protect Neurons against Ischemia by HIF-1α–Regulated VEGF Signaling

doi: 10.1371/journal.pone.0009767

Figure Lengend Snippet: ( A ) Diagram of transwell coculture. Micrograph depicts cortical neurons in lower compartment immunofluorescently labeled for MAP-2 (red) and GFAP (green). NSPCs in upper compartment are immunofluorescent for nestin (red). Nuclei are labeled with DAPI (blue). ( B ) Neuronal survival at 24 hrs following 2 hr exposure to OGD in the absence (left) or presence (right) of NSPCs. ( C ) Survival of NSPCs grown as monocultures in upper compartment. Data were acquired using the MTT colorimetric viability assay as described in section. *p<0.01, Student's t-test, n = 4 cultures/group.

Article Snippet: Cultured cells were fixed on coverslips with 4% paraformaldehyde and incubated in monoclonal mouse anti-Nestin (1∶500; Chemicon, Temecula, CA, USA), polyclonal rabbit anti-MAP-2 (1∶500; Chemicon), and/or polyclonal rabbit anti-GFAP (1∶500; Accurate, Westbury, NY, USA).

Techniques: Labeling, Viability Assay

Journal: PLoS ONE

Article Title: Granulocyte-Colony Stimulating Factor Attenuates Oligomeric Amyloid β Neurotoxicity by Activation of Neprilysin

doi: 10.1371/journal.pone.0103458

Figure Lengend Snippet: A–E, Fluorescent microscopic images of mouse primary cortical neuron cultures. A, Untreated neurons. B, Neurons treated with 5 µM oAβ 1–42 . C, Neurons treated with 5 µM oAβ 1–42 and 1 ng/ml G-CSF. D, Neurons treated with 5 µM oAβ 1–42 and 10 ng/ml G-CSF. E, Neurons treated with 5 µM oAβ 1–42 and 100 ng/ml G-CSF. Treatment with G-CSF was neuroprotective against oAβ-mediated toxicity. Neurons were stained with anti–MAP-2 antibodies (green), and Aβ was stained with 4G8 antibodies (red). Scale bar: 50 µm. F, Relative neuronal survival. The number of viable neurons (MAP-2–positive neurons) was quantified relative to results observed with untreated neurons. G-CSF rescued neurons against oAβ-mediated toxicity in a dose-dependent manner. *, p <0.001; †, p <0.01; ††, p <0.05. Values are means ± SEM (n = 3). G, WST-8 assay. G-CSF enhanced neuronal survival against oAβ-mediated toxicity in a dose-dependent manner. *, p <0.001; †, p <0.01; ††, p <0.05. Values are means ± SEM (n = 6).

Article Snippet: After blocking with 5% goat serum for 1 h at room temperature, cells were stained with rabbit polyclonal anti-microtubule–associated protein (MAP)-2 antibody (1∶1000, Millipore, Billerica, MA, USA), and Aβ was stained with mouse monoclonal anti-Aβ antibody (clone 4G8, 1∶1000, Millipore).

Techniques: Staining

Journal: PLoS ONE

Article Title: Granulocyte-Colony Stimulating Factor Attenuates Oligomeric Amyloid β Neurotoxicity by Activation of Neprilysin

doi: 10.1371/journal.pone.0103458

Figure Lengend Snippet: A–F, Fluorescent microscopic images of mouse primary cortical neuron cultures. A, Untreated neurons. B, Neurons treated with 5 µM oAβ 1–42 . C, Neurons treated with 5 µM oAβ 1–42 and 100 ng/ml G-CSF. D, Neurons treated with 5 µM oAβ 1–42 , 100 ng/ml G-CSF and 0.1 ng/ml thiorphan. E, Neurons treated with 5 µM oAβ 1–42 , 100 ng/ml G-CSF and 1 ng/ml thiorphan. F, Neurons treated with 5 µM oAβ 1–42 , 100 ng/ml G-CSF and 10 ng/ml thiorphan. The NEP inhibitor canceled the neuroprotective effects of G-CSF. Neurons were stained with anti–MAP-2 antibodies (green) and Aβ was stained with 4G8 antibodies (red). Scale bar: 50 µm. G, Relative neuronal survival. The number of viable neurons (MAP-2–positive neurons) was quantified relative to results observed with untreated neurons. The NEP inhibitor dose-dependently suppressed the neuroprotective effects of G-CSF. *, p <0.001; †, p <0.001; ‡, p <0.05; §, p <0.01. Values are means ± SEM (n = 3). H, WST-8 assay. The NEP inhibitor reversed the neuroprotective effects of G-CSF in a dose-dependent manner. *, p <0.001; †, p <0.001; ‡, p <0.05; §, p <0.01. Values are means ± SEM (n = 6).

Article Snippet: After blocking with 5% goat serum for 1 h at room temperature, cells were stained with rabbit polyclonal anti-microtubule–associated protein (MAP)-2 antibody (1∶1000, Millipore, Billerica, MA, USA), and Aβ was stained with mouse monoclonal anti-Aβ antibody (clone 4G8, 1∶1000, Millipore).

Techniques: Staining

Journal: PLoS ONE

Article Title: Granulocyte-Colony Stimulating Factor Attenuates Oligomeric Amyloid β Neurotoxicity by Activation of Neprilysin

doi: 10.1371/journal.pone.0103458

Figure Lengend Snippet: A–F, Fluorescent microscopic images of mouse primary cortical neuron cultures. A, Untreated neurons. B, Neurons treated with 5 µM oAβ 1–42 . C, Neurons treated with 5 µM oAβ 1–42 and 100 ng/ml G-CSF. D, Neurons treated with 5 µM oAβ 1–42 , 100 ng/ml G-CSF and 0.3 µM BIX02189. E, Neurons treated with 5 µM oAβ 1–42 , 100 ng/ml G-CSF and 3 µM BIX02189. F, Neurons treated with 5 µM oAβ 1–42 , 100 ng/ml G-CSF and 30 µM BIX02189. The MEK5/ERK5 inhibitor BIX02189 almost completely suppressed G-CSF–mediated protection against oAβ-induced neurotoxicity. Neurons were stained with anti–MAP-2 antibodies (green), and Aβ was stained with 4G8 antibodies (red). Scale bar: 50 µm. G, Relative neuronal survival. The number of viable neurons (MAP-2–positive neurons) was quantified relative to results observed with untreated neurons. Inhibition of MEK5/ERK5 almost completely ablated the neuroprotective effects of G-CSF. *, p <0.001; †, p <0.001; ‡, p <0.001. Values are means ± SEM (n = 3). H, WST-8 assay. Blocking MEK5/ERK5 almost completely canceled the neuroprotective effects of G-CSF. *, p <0.001; †, p <0.001; ‡, p <0.001. Values are means ± SEM (n = 6).

Article Snippet: After blocking with 5% goat serum for 1 h at room temperature, cells were stained with rabbit polyclonal anti-microtubule–associated protein (MAP)-2 antibody (1∶1000, Millipore, Billerica, MA, USA), and Aβ was stained with mouse monoclonal anti-Aβ antibody (clone 4G8, 1∶1000, Millipore).

Techniques: Staining, Inhibition, Blocking Assay

Journal: PLoS ONE

Article Title: A Defined and Xeno-Free Culture Method Enabling the Establishment of Clinical-Grade Human Embryonic, Induced Pluripotent and Adipose Stem Cells

doi: 10.1371/journal.pone.0010246

Figure Lengend Snippet: A ) RT-PCR analysis of in vitro -derived EBs showing transcripts for AFP and SOX17 (endodermal markers), α-cardiac actin and T (Brachyury; mesodermal markers), SOX1 and PAX6 (ectodermal markers), and β-actin . Lane 1, 50-bp DNA ladder. Regea 07/046 at passage 42, Regea 08/013 at passage 35, and Regea 06/040 at passage 101. B ) Differentiated cardiomyocytes from hESC line Regea 08/013 were stained with a) cardiac troponin T (red) and b) ventricular myosin heavy chain (green). c) Merged picture of a) and b) with DAPI staining. Scale bar is 100 µm. C ) RT-PCR analysis of neurospheres derived from hESC line Regea 08/013 cultured in RegES medium showed expression of neural precursor markers Musashi , Nestin and PAX6 ; neuronal markers MAP-2 , NF68 and OTX2 ; and astrocytic marker GFAP . No expression of pluripotent markers Oct4 and Nanog , nor endo- AFP or mesodermal markers T/Brachyury were detected. D ) Most of the cells migrating out from the plated neurospheres stained positive for neuronal marker MAP-2 (green) and few cells were positive for astrocytic marker GFAP (red). DAPI staining (blue). Scale bar is 100 µm.

Article Snippet: The fixed neuronal cells were incubated overnight at 4°C with polyclonal rabbit anti- MAP-2 (Chemicon) for neuronal cells and polyclonal sheep anti-GFAP (R&D Systems) for astrocytes.

Techniques: Reverse Transcription Polymerase Chain Reaction, In Vitro, Derivative Assay, Staining, Cell Culture, Expressing, Marker

Journal: Molecular Pain

Article Title: Ultra-low dose naltrexone attenuates chronic morphine-induced gliosis in rats

doi: 10.1186/1744-8069-6-22

Figure Lengend Snippet: The morphine-induced increase in astrocyte and microglial immuno-labelling is caused by cell hypertrophy, not proliferation . Lumbar spinal cord sections were collected from animals administered BrdU (100 mg/kg) by intraperitoneal injection on days 1, 3, 5, and either intrathecal vehicle (saline; SAL) or morphine (15 μg; MS) once daily for five days by lumbar puncture. Representative photomicrographs acquired by confocal microscopy of spinal cord sections double labelled with 5-bromo-2-deoxyuridine (BrdU) and the astrocytic protein GFAP (B, C), the microglial marker Iba1 (E, F) or the neuronal marker MAP2 (H, I). No co-localization of BrdU-positive cells with GFAP or MAP-2-positive cells was observed. However, BrdU co-localized with a small number of Iba1 positive cells (arrow), suggesting a small portion of the newly formed cells were microglia or macrophages. (J). No difference was observed in the number of BrdU-positive cells in the dorsal horn (lamina II-IV) of lumbar spinal cord sections from animals administered chronic intrathecal saline or morphine. Data represent means ± s.e.m. for n = 6 sections per rat from n = 3 per group. Statistical analyses were performed by an un-paired t-test. ns = no significance. Scale bars, 30 μm.

Article Snippet: To identify the phenotype of newly formed cells, sections were double labelled with one of the following antibodies: rabbit anti-GFAP (for astrocytes, 1:2500), rabbit anti-Iba1 polyclonal antibody (ionizing calcium-binding adaptor molecule, for microglia and macrophages, 1:1000; Wako, Richmond, VA), or rabbit anti-MAP-2 polyclonal antibody (microtubule-associated protein 2, for neurons, 1:1000; Chemicon, Temecula, CA).

Techniques: Injection, Confocal Microscopy, Marker