Journal: Neural Plasticity

Article Title: Transplantation of Neural Stem Cells Cotreated with Thyroid Hormone and GDNF Gene Induces Neuroprotection in Rats of Chronic Experimental Allergic Encephalomyelitis

doi: 10.1155/2016/3081939

Figure Lengend Snippet: Immunohistochemical double staining of BrdU, MAP2, GFAP, and GalC positive cells in the cerebral parenchyma of rats. (a–c) MAP2 − /BrdU + cells (solid arrow), MAP2 + /BrdU − cells (dotted arrow), and MAP2 + /BrdU + cells (arrowhead) in NSCs, T3/NSCs, and GDNF-T3/NSCs group, respectively. (d–f) GalC − /BrdU + positive cells (solid arrow), GalC + /BrdU − cells (dotted arrow), and GalC + /BrdU + cells (arrowhead) in NSCs, T3/NSCs, and GDNF-T3/NSCs group, respectively. (g–i) GFAP − /BrdU + cells (solid arrow), GFAP + /BrdU − cells (dotted arrow), and GFAP + /BrdU + cells (arrowhead) in NSCs, T3/NSCs, and GDNF-T3/NSCs group, respectively. (j) GFAP + cells (dotted arrow) in control group. (k) GFAP + cells (dotted arrow) in normal group. Bar (a–k) = 25 μ m.

Article Snippet: After a goat anti-mouse IgG secondary antibody (1 : 100; Wuhan Boster Biological Technology, China) was added for 30 minutes, alkaline phosphatase- (AP-) streptavidin (Boster) was incubated for 30 minutes, and 5-bromo-4-chloro-3-indolyl phosphate/nitroblue tetrazolium chloride (BCIP/NBT, Boster) was then used as a chromogen for 10 min. After PBS washes, the sections were reincubated with primary antibody for rabbit ant-MAP2, GFAP, or GalC, followed by incubation with goat anti-rabbit IgG (1 : 100; Boster) and horseradish peroxidase- (HRP-) streptavidin (Boster).

Techniques: Immunohistochemical staining, Double Staining

Journal: Neural Plasticity

Article Title: Transplantation of Neural Stem Cells Cotreated with Thyroid Hormone and GDNF Gene Induces Neuroprotection in Rats of Chronic Experimental Allergic Encephalomyelitis

doi: 10.1155/2016/3081939

Figure Lengend Snippet: Differentiation of NSCs, T3/NSCs, and GDNF-T3/NSCs in vivo (mean ± SD, n = 5).

Article Snippet: After a goat anti-mouse IgG secondary antibody (1 : 100; Wuhan Boster Biological Technology, China) was added for 30 minutes, alkaline phosphatase- (AP-) streptavidin (Boster) was incubated for 30 minutes, and 5-bromo-4-chloro-3-indolyl phosphate/nitroblue tetrazolium chloride (BCIP/NBT, Boster) was then used as a chromogen for 10 min. After PBS washes, the sections were reincubated with primary antibody for rabbit ant-MAP2, GFAP, or GalC, followed by incubation with goat anti-rabbit IgG (1 : 100; Boster) and horseradish peroxidase- (HRP-) streptavidin (Boster).

Techniques: In Vivo

Journal: Oxidative Medicine and Cellular Longevity

Article Title: The Role of the miR-548au-3p/CA12 Axis in Tracheal Chondrogenesis in Congenital Pulmonary Airway Malformations

doi: 10.1155/2023/6428579

Figure Lengend Snippet: Identification of DEGs involved in the pathogenesis of CPAM. (a) Flowchart of the study design and samples at each stage of analysis. (b) Scatterplot of mRNA expression variation between diseased CPAM and normal tissues. (c) Hierarchical cluster of gene expression profiles from microarray assays. Expression of CA12, LONRF3, MAP2, THBS1, and PPID at mRNA (d) and protein (e) levels in lung tissues from CPAM and adjacent normal tissues. ∗∗∗ p < 0.001, compared with normal.

Article Snippet: Primary antibodies for this study were specific for CA12 (Boster, Cat. No. A04063, 1 : 1000), LONRF3 (GeneTex, Cat. No. GTX112150, 1 : 2000), MAP2 (Boster, Cat. No. A01201, 1 : 2000), THBS1 (Boster, Cat. No. PB0471, 1 : 2000), PPID, E-cadherin (Beyotime, Cat. No. AF6759, 1 : 1000), N-cadherin (Beyotime, Cat. No. AF5237, 1 : 800), aggrecan (Abcam, Cat. No. ab3778, 1 : 1000), Col2A1 (Boster, Cat. No. A00517, 1 : 2000), MMP13 (Proteintech, Cat. No. 18165-1-AP, 1 : 3000), ADAMTS4 (Proteintech, Cat. No. 11865-1-AP, 1 : 600), and GAPDH (Abcam, Cat. No. ab9485, 1 : 2000).

Techniques: Expressing, Gene Expression, Microarray

Journal: Cell Death & Disease

Article Title: Human iNSC-derived brain organoid model of lysosomal storage disorder in Niemann–Pick disease type C

doi: 10.1038/s41419-020-03262-7

Figure Lengend Snippet: a Schematic of the method for the generation of WT organoids using iNSCs. b Bright-field images of the organoids at day 28 of maturation. Scale bars, 500 μm. c Quantification of the diameter (mm) during the entire course of the brain organoid development under different culture conditions. The results are the mean ± SD. *** p < 0.001. n = 10 diameters of organoids were quantified per time point. d RT-qPCR analysis of neural progenitor cell markers, SOX2 and PAX6 , an intermediate progenitor marker, TBR2 , and neuron markers, TUJ1, MAP2 , and NF , in the organoids at day 28. The data are normalized to iNSCs at day 0. The results are the mean ± SEM. n = 3 per sample. e Clearing and immunohistochemistry revealed the whole-mount morphology with heterogeneous regions containing neural progenitors (SOX2, red) and neurons (TUJ1, green). Scale bars, 500 μm. Zoom scale bars, 60 μm. f Immunohistochemical staining of serial sections for neuronal markers (TUJ1, MAP2, and NF) at day 28 of differentiation. Scale bars, 50 μm.

Article Snippet: The primary antibodies used were TUJ1 (1:1000, Biolegend, 801202), MAP2 (1:500, Merck, MAB3418), NF (1:1000, Cell Signaling, 2836s), ß-actin (1:1000, CST, 4967), cleaved-caspase-3 (1:1000, CST, 9664S), Caspase-3 (1:1000, CST, 9662), LC3 (1:1000, Novus, NB100), and p62 (1:500, BD Bioscience, 610832).

Techniques: Quantitative RT-PCR, Marker, Immunohistochemistry, Immunohistochemical staining, Staining

Journal: Cell Death & Disease

Article Title: Human iNSC-derived brain organoid model of lysosomal storage disorder in Niemann–Pick disease type C

doi: 10.1038/s41419-020-03262-7

Figure Lengend Snippet: a Clearing and immunostaining with DAPI in the WT and NPC organoids at day 28. Scale bars, 500 μm. b Immunostaining for mitotic radial glia (phospho-vimentin) in the outer region. Scale bars, 50 μm. c Staining for neuronal markers (TUJ1, NF, and MAP2) in the WT and NPC organoids at day 28. Scale bars, 50 μm. d The percentage of the total number of cells that expressed TUJ1, NF, and MAP2 of the total number of DAPI-positive cells counted in each group was determined. e , f Western blot analysis and quantification of neuronal markers (TUJ1, NF, and MAP2) reveals a reduction in the NPC organoids on day 28. Every experiment was performed in triplicate. The results are the mean ± SD. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: The primary antibodies used were TUJ1 (1:1000, Biolegend, 801202), MAP2 (1:500, Merck, MAB3418), NF (1:1000, Cell Signaling, 2836s), ß-actin (1:1000, CST, 4967), cleaved-caspase-3 (1:1000, CST, 9664S), Caspase-3 (1:1000, CST, 9662), LC3 (1:1000, Novus, NB100), and p62 (1:500, BD Bioscience, 610832).

Techniques: Immunostaining, Staining, Western Blot

Journal: Frontiers in Neuroanatomy

Article Title: Analysis of the expression and distribution of protein O-linked mannose β1,2- N -acetylglucosaminyltransferase 1 in the normal adult mouse brain

doi: 10.3389/fnana.2022.1043924

Figure Lengend Snippet: Primary antibodies used in this work.

Article Snippet: MAP2 , Mouse, mAb , Signalway, Shanghai, China , — , — , 1:200.

Techniques: Immunohistochemistry-IF

Journal: Frontiers in Neuroanatomy

Article Title: Analysis of the expression and distribution of protein O-linked mannose β1,2- N -acetylglucosaminyltransferase 1 in the normal adult mouse brain

doi: 10.3389/fnana.2022.1043924

Figure Lengend Snippet: Cellular localization of POMGNT1. (A–I) Dual immunostaining of POMGNT1 (red) and neuronal glial cell subtype-specific markers (green) in the coronal brain sections of the cerebral cortex, including MAP2 for mature neurons (A) , S100B for resting and activated astrocytes (B) , GFAP for activated astrocytes (C) , MBP for oligodendrocytes (D) , or Iba-1 for microglia (E) , VGLUT1 for Glutamatergic neurons (F) , and GAD65 for GABAergic neurons (G) . POMGNT1 expressed in some neuronal glial cell subtype-specific marker-positive cells, including MAP2, S100B, MBP, Iba-1, GAD65, and VGLUT1, suggesting POMGNT1 may distribute in mature neurons, astrocytes, oligodendrocytes, microglia, glutamatergic neurons, and GABAergic neurons. Nearly no POMGNT1 expressed in the GFAP-positive cells suggesting POMGNT1 may not distribute in the activated astrocytes. N = 6. Scale bars = 20 μM. (H) Quantification of the percentage of the marker labeling cells containing POMGNT1 shown in panels (A–G) . N = 6. Data are presented as the mean ± SEM; ** P < 0.01; *** P < 0.001 versus the MAP2 group (one-way ANOVA); # P < 0.05 versus the VGLUT1 group (Student’s t -test). (I) The protein expression of POMGNT1 in neurons or glial cells using in vitro culturing, including HT22, MA-c, MOPC, and BV2, was detected by Western blot. β-actin as the loading control. (J) The results of the western blot were quantified. N = 4. Data are presented as the mean ± SEM; ** P < 0.01; **** P < 0.0001 versus the HT22 group (one-way ANOVA).

Article Snippet: MAP2 , Mouse, mAb , Signalway, Shanghai, China , — , — , 1:200.

Techniques: Immunostaining, Marker, Labeling, Expressing, In Vitro, Western Blot, Control

Journal: Cancer Medicine

Article Title: microRNA‐196a‐5p inhibits testicular germ cell tumor progression via NR6A1/E‐cadherin axis

doi: 10.1002/cam4.3498

Figure Lengend Snippet: NR6A1 promoted neural cell‐like characteristics of testicular tumor in vivo and in vitro. A, At the experimental end point, tumor xenografts were dissected and photographed. B, Tumor weights were measured at the experimental end point, ** p < 0.01. C, Tumor volume was measured every 3 days after injection of tumor cells, * p < 0.05. D, Immunohistochemistry staining of NR6A1, PCNA, E‐cadherin, and MAP2 protein in NT‐2 testicular tumor xenografts. E, Diagram for establishment of RA‐induced NT‐2 cell neurodifferentiation model in vitro. F, The morphological changes in NT‐2 cells under different RA induction days. G, Immunofluorescence analysis of the expression and localization of MAP2 protein in NT‐2 cells with RA induction of 21 days. H, RT‐PCR analysis of MAP2 in long‐term RA‐induced NT‐2 cells with NR6A1 interference. I, Western blot analysis of MAP2 in long‐term RA‐induced NT‐2 cells with NR6A1 interference. All data are shown as the means ±SDs of three independent experiments

Article Snippet: Cells were fixed with 4% paraformaldehyde (Sigma) for 15 min and permeated with 0.2% Triton X‐100 for 5 min. After washed with PBS buffer three times for 5 min, cells were blocked with fetal bovine serum for 30 min, then incubated with the primary antibody at 4°C overnight [E‐cadherin (1:100, CST, #3195); MAP2 (1:40, Sangon Biotech, AF2215)].

Techniques: In Vivo, In Vitro, Injection, Immunohistochemistry, Staining, Immunofluorescence, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: Cancer Medicine

Article Title: microRNA‐196a‐5p inhibits testicular germ cell tumor progression via NR6A1/E‐cadherin axis

doi: 10.1002/cam4.3498

Figure Lengend Snippet: The correlation of miR‐196a‐5p/NR6A1/E‐cadherin in RA‐induced NT‐2 cells and clinical TGCTs tissue specimens. A, Western blot analysis of NR6A1, OCT4, E‐cadherin, and N‐cadherin expression in NT‐2 cells treated with different concentrations of RA for 24 h. B, Western blot analysis of NR6A1, OCT4, E‐cadherin, and N‐cadherin expression in NT‐2 cells treated with 10 μmol/L RA for different numbers of days. C, RT‐qPCR detection of miR‐196a‐5p expression in RA‐treated NCCIT and NT‐2 cells. ** represented p < 0.01 compared with cells without RA treatment group. D, The expression alteration of miR‐196a‐5p/NR6A1/E‐cadherin in NT‐2 cells with 10 μmol/L RA treatment for 0‐5 days by RT‐qPCR. * represented p < 0.05 compared with RA treated 0 day group; # represented p < 0.05 compared with NC mimics group without RA treatment. E, Immunohistochemistry analysis of NR6A1, E‐cadherin, and MAP2 expression in both normal testes and TGCTs. Up panel showed the immunohistochemistry staining of NR6A1, E‐cadherin, and MAP2 protein in both normal testes and TGCTs. Down panel showed the immunohistochemistry scores of NR6A1, E‐cadherin, and MAP2 in both normal testes and TGCTs (n = 15). ** p < 0.01. SEM, seminoma. EC, testicular embryonal carcinomas. (F) The correlation between NR6A1 and E‐cadherin (r = −0.6576, p = 0.0077), NR6A1 and MAP2 ( r = 0.5221, p = 0.0459), E‐cadherin and MAP2 ( r = −0.0599, p = 0.8321), was analyzed based on immunohistochemistry staining score in clinical TGCTs. The correlation is deemed significant and positive when p < 0.05. All data are shown as the means ±SDs of three independent experiments

Article Snippet: Cells were fixed with 4% paraformaldehyde (Sigma) for 15 min and permeated with 0.2% Triton X‐100 for 5 min. After washed with PBS buffer three times for 5 min, cells were blocked with fetal bovine serum for 30 min, then incubated with the primary antibody at 4°C overnight [E‐cadherin (1:100, CST, #3195); MAP2 (1:40, Sangon Biotech, AF2215)].

Techniques: Western Blot, Expressing, Quantitative RT-PCR, Immunohistochemistry, Staining