primary polyclonal rabbit anti-map2 antibody  (Millipore)

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: PLoS Genetics

Article Title: FOXR1 regulates stress response pathways and is necessary for proper brain development

doi: 10.1371/journal.pgen.1009854

Figure Lengend Snippet: (A) Representative schematic view of the CRISPR/Cas9 targeting strategy used for generating Foxr1 knockout mice. The Foxr1 locus consists of 5 exons (colored boxes), and introns (black lines). Two gRNAs (red lines), one upstream located in intron 1 and one downstream located within exon 4 were used to target the Foxr1 gene and remove 976 nucleotides encompassing exons 2, 3 and part of exon 4 (blue box). Primers are represented by black half arrowheads to indicate the relative locations of forward (primer 1) and reverse genotyping primers (primers 2 and 3). (B) Representative PCR genotyping result for primer set 1 and 2 (indicated in panel A) to detect Foxr1 wild-type allele (354 bp). Below is a representative PCR genotyping result for primer set 1 and 3 to detect Foxr1 wild-type (1381 bp), heterozygous (1381 for wild-type and 403 bp for knockout), and knockout alleles (403 bp). (C) RT-PCR of Foxr1 (175 bp) and 18S ribosomal transcripts (129 bp) from brains of wild-type and Foxr1 knockout mice. (D) Genotype analysis of number of offspring obtained from Foxr1 heterozygous crossings at postnatal day 21 (P21). (E) Genotype analysis of number of newborn offspring obtained from Foxr1 heterozygous crossings at postnatal day 0 (P0). (F) Lateral view of wild-type and Foxr1 knockout neonates showing a decrease in size in the Foxr1 knockout mutant. (G) Body weight measurements (in grams) for Foxr1 wild-type (n = 53), heterozygous (n = 120) and knockout mice (n = 12) at postnatal day 0 showing a decrease in Foxr1 knockout mice. One-way ANOVA Tukey’s multiple comparisons (*** p < 0.001). (H) Nissl stain brain sections of Foxr1 wild-type (+/+), heterozygous (+/-) and knockout (-/-) mice. Top panel shows coronal sections anterior to bregma. Dashed lines represent cortical measurements at 0°, 45° and 90° (relative to the midline) to pia surface. White box indicates higher magnification for the bottom panels. Scale bar = 500 μm. Bottom panels represents higher magnification of the cortex where the vertical line indicates cortical thickness measurements. Side bars demarcates the different cortical layers. Scale bar = 125 μm. (I) MAP2 immunostaining of brain sections of Foxr1 wild-type and knockout mice. Top panel shows coronal sections anterior to bregma. White box indicates higher magnification for the bottom panels. Scale bar = 500 μm. Bottom panels represents higher magnification of the cortex where the vertical line indicates cortical thickness measurements. Scale bar = 125 μm. (J-L) Quantification of cortical thickness from pooled brain sections of 4 wild-type, 4 heterozygous and 4 Foxr1 knockout mice at 0°, 45° and 90° (relative to the midline) to pia surface, respectively. Graph represents relative thickness normalized to wild-type (WT). One-way ANOVA Tukey’s multiple comparisons (0°, * p = 0.0165; 45°, ** p = 0.0033, *** p = 0.0003; 90°, ** p = 0.0019, *** p <0.0001) (M) Quantification of ventricle area from pooled brain sections of 4 wild-type and 4 Foxr1 knockout mice. Graph represents relative thickness normalized to wild-type (WT). One-way ANOVA Tukey’s multiple comparisons * p = 0.04 wild-type and knockout; p = 0.02 heterozygous and knockout.

Article Snippet: Brain sections were then incubated with primary polyclonal rabbit anti-MAP2 antibody (1:500 dilution in 5% goat serum in PBS; cat# AB5622 Millipore) at 4°C overnight.

Techniques: CRISPR, Knock-Out, Reverse Transcription Polymerase Chain Reaction, Mutagenesis, Staining, Immunostaining

Journal: Cells

Article Title: Vertically-Aligned Functionalized Silicon Micropillars for 3D Culture of Human Pluripotent Stem Cell-Derived Cortical Progenitors

doi: 10.3390/cells9010088

Figure Lengend Snippet: hCPs seeded on silicon micropillar arrays preserve their cortical regional identity and upon exposure to differentiative conditions, generate cortical glutamatergic neurons. ( A ) eGFP +ve hCPs cultured for 14 days on 3D silicon devices preserve the expression of the cortical progenitor marker TBR2 (red). Nuclei are stained with Hoechst (blue). Scale bar: 50 μm. ( B ) hCPs cultured for 5 days on 3D silicon devices and then exposed for 35 days to differentiative conditions maturate into cortical glutamatergic neurons. Left: cultures stained for the pan-neuronal neuronal marker β3-TUBULIN (red). Nuclei are stained with Hoechst (blue). Scale bar: 10 μm. Right: cultures stained for the mature neuronal marker MAP2 (red) and for the cortical neuronal marker CUX1 (green). Nuclei are stained with Hoechst (blue). Scale bar: 10 μm.

Article Snippet: Antibodies used in this study: primary mouse monoclonal anti-NESTIN antibody (R&D Systems, Minneapolis, MN, USA, 1:300), primary mouse monoclonal β3-TUBULIN antibody (Promega, Milan, Italy, 1:1000), primary rabbit polyclonal anti-SOX2 antibody (Millipore, Milan, Italy, 1:200), primary rabbit polyclonal anti-MAP2 antibody (Santa Cruz, Heidelberg, Germany, 1:200), primary mouse monoclonal anti-TBR2 antibody (ABCAM, Cambridge, UK, 1:500), primary mouse monoclonal anti CUX1 (ABCAM, 1:200), AlexaFluor-488 or -568 conjugated secondary antibodies (Thermo Fisher Scientific, 1:500).

Techniques: Cell Culture, Expressing, Marker, Staining