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: Neurobiology of disease

Article Title: Cypin: A novel target for traumatic brain injury.

doi: 10.1016/j.nbd.2018.07.019

Figure Lengend Snippet: (a) Representative images showing neurons immunostained for the neuronal marker, MAP2 and GFP and co-stained with nuclear dye, Hoechst 33225, after treatment with vehicle (0.1% DMSO; control), 20 μM NMDA, with or without 48h pretreatment with 20μM H9, B9, G5, G6. Scale bar = 100μm. (b-g). Quantitative analysis of neuronal survival expressed as percent live control neurons. Cypin activators, H9 and G5, prevent neuronal loss after injury, while the inhibitor, B9, and the neutral compound, G6, have no beneficial effect in control cultures. Cypin knockdown induces 75% cell death, and abolishes neuroprotective effects of H9 and G5. Data represent 39-43 samples from three separate trials. **p<0.01, ***p<0.005, ****p<0.001 determined by one-way ANOVA followed by Tukey-Kramer multiple comparisons test. Error bars indicate ± SEM. (h) Representative images showing neurons immunostained for the neuronal marker, MAP2 and GFP, and co-stained with nuclear dye, Hoechst 33225, after treatment with vehicle (0.1% DMSO; control), 20 μM NMDA, with or without 48h pretreatment with 20 μM H9, B9, G5, G6. Scale bar = 100μm. (i-l). Quantitative analysis of neuronal survival expressed as percent live control neurons. Cypin activators, H9 and G5, prevent neuronal loss after injury, while the inhibitor, B9, and the neutral compound, G6, have no beneficial effect in control cultures. Cypin overexpression prevents neuronal death, and this effect is abolished by pretreatment with B9. Data represent 32-44 samples from three separate trials. *p<0.05, **p<0.01, ***p<0.005, ****p<0.001 determined by one-way ANOVA followed by Tukey-Kramer multiple comparisons test. Error bars indicate ± SEM.

Article Snippet: Cultures were fixed in 4% paraformaldehyde in phosphate-buffered saline (PBS) for 10 min, permeabilized with 0.1% Triton X-100 in PBS + 5% normal goat serum, and immunostained with mouse anti-MAP2 (1:500) from Rockland and anti-GFP (1:500) from EMD Millipore followed by secondary antibodies conjugated to Alexa-Fluor® 488 (Invitrogen, 1:250) or Alexa-Fluor® 555 (Invitrogen, 1:250).

Techniques: Marker, Staining, Over Expression

Journal: Neuron

Article Title: Targeted proteoform mapping uncovers specific Neurexin-3 variants required for dendritic inhibition

doi: 10.1016/j.neuron.2022.04.017

Figure Lengend Snippet:

Article Snippet: The following antibodies were used in this study: rabbit polyclonal anti-β-actin (Abcam; Cat# ab8227; RRID: AB_2305186; LOT# GR3314266-1), mouse monoclonal anti-calbindin (Swant; Cat# 300; RRID: AB_10000347; LOT# 17 (F)), goat polyclonal anti-calretinin (Swant; Cat# CG1; RRID: AB_10000342; LOT# 1§.1), mouse monoclonal anti-CamKII alpha (Thermo Fisher Scientific; 6G9; Cat# Ma1-048; RRID: AB_325403; LOT# TH269517), mouse monoclonal anti-cannabinoid receptor 1 (Immunogene; IMG-3C2; Cat# IMG-CB1R-mAb001; LOT# CJ03), guinea pig polyclonal anti-cholecystokinin (Synaptic Systems, Cat# 438004; RRID: AB_2814938; LOT# 1-1), mouse monoclonal anti-GAD67 (Millipore; 1G10.2; Cat# MAB5406; RRID: AB_2278725; LOT# 3015328), rabbit polyclonal anti-GAPDH (Enogene; Cat# E1C604; LOT# R14Q12), mouse monoclonal anti-gephyrin (Synaptic Systems; mAb7a; Cat# 147021; RRID: AB_2232546; LOT# 147021/15), mouse monoclonal anti-gephyrin (Synaptic Systems; mAb7a; Cat# 147011; RRID: AB_887717; LOT# 147011/54), rat monoclonal anti-HA (Roche; 3F10; Cat# 11867431001; RRID: AB_390919; LOT# 34502100), rabbit monoclonal anti-HA (Cell Signaling; 3724; Cat# 3724; RRID: AB_1549585; LOT# 9), guinea pig polyclonal anti-MAP2 (Synaptic Systems; Cat# 188004; RRID: AB_2138181; LOT# 2-26), mouse monoclonal anti-MAP2 (Synaptic Systems; 198A5; Cat# 188011; RRID: AB_2147096; LOT# 1-10), rabbit polyclonal anti-neurexin , chicken anti-neurexin , rabbit anti-neuroligin , rabbit monoclonal anti-nNOS (Cell Signaling; C7D7; Cat# 4231; RRID: AB_2152485; LOT# 2), goat polyclonal anti-parvalbumin (Swant, Cat# PVG214; RRID: AB_10000345), mouse monoclonal anti-PSD95 (Santa Cruz; 7E3; Cat# sc32290; RRID: AB_628114; LOT# J1509), goat polyclonal anti-somatostatin (Santa Cruz; Cat# sc7819; RRID: AB_2302603; LOT# L1611), mouse monoclonal anti-Synaptotagmin 2 (Zebrafish International Resource Center; Cat# znp-1; RRID: AB_10013783), mouse monoclonal anti-V5 (Biorad; SV5-PK1; Cat# MCA1360; RRID: AB_322378; LOT# 148239), guinea pig polyclonal anti-vGAT (Synaptic Systems; Cat# 131004; RRID: AB_887873; LOT# 2-42), guinea pig polyclonal anti-vGlut1 (Millipore; Cat# AB5905; RRID: AB_2301751; LOT# 3308226), rabbit polyclonal anti-VIP (Immunostar; Cat# 20077; RRID: AB_572270; LOT# 1513001), guinea pig anti-somatostatin (raised in the present study against amino acid residues 35-88 of mouse pro-somatostatin, GenBank: #BC010770.1), goat anti-cannabinoid receptor 1 (Nittobo Medical, MSFR100600; RRID: AB_2571592), guinea pig anti-GABAA receptor alpha 1 (Nittobo Medical, MSFR101540; RRID: AB_2571572), goat anti-vGAT (Nittobo Medical, MSFR106130; RRID: AB_2571623), guinea pig anti-PSD95 (Nittobo Medical, MSFR105180; RRID: AB_2571612).

Techniques: Recombinant, Reporter Assay, Multiplex Assay, Plasmid Preparation, Software, Microscopy