Journal: Nature communications

Article Title: A neurodegeneration checkpoint mediated by REST protects against the onset of Alzheimer's disease.

doi: 10.1038/s41467-023-42704-6

Figure Lengend Snippet: Fig. 3 | Role of the UPR and Wnt/β-catenin signaling in nuclear REST induction. a Immunolabeling of UPR activation (marker BiP/GRP78, red), REST (green), β- catenin (magenta) and DNA (DAPI, blue) in 11-month-old 3xTg and WT mice shows coordinate upregulation of BiP, nuclear REST and nuclear β-catenin expression in 3xTg mice. b Correlation between nuclear β-catenin and nuclear REST (left graph), BiP and nuclear REST (middle graph) and BiP and nuclear β-catenin (right graph) levels in the hippocampus CA1 region of 11-month-old 3xTg mice. Shown are the mean fluorescence intensity values for nuclear REST in individual CA1 neurons from n = 3 3xTg mice. a.u.- arbitrary units. The Pearson correlation coefficient (r) and P- value are shown. c Immunolabeling of REST (green) and neuron marker MAP2 (red) in 3xTg primary cortical neurons (PCNs) shows nuclear REST in neurons treated with vehicle and decreased nuclear REST in 3xTg neurons after a 24-h treatment with the Wnt/β-catenin inhibitors Dickkopf 1 (DKK1), XAV939 and ICG001, or the PERK inhibitor GSK2606414. d Quantification of average nuclear REST levels in WT PCNs, as well as 3xTg PCNs treated with vehicle or the individual drugs. The P-

Article Snippet: Additional primary antibodies were as follows: anti-human Aβ rabbit monoclonal IgG antibody (Cell Signaling, Cat. No. 8243); antihuman APP mouse monoclonal IgG antibody (clone 6E10; Covance, Catalog No. SIG-39320); anti-actin mouse monoclonal IgG antibody (clone ACTN05 (C4); ThermoFisher Scientific, CatalogNo.MA5-11869); anti-NeuN mouse monoclonal IgG antibody (clone A60, Millipore, MAB377); anti-MAP2 goat polyclonal IgG antibody (PhosphoSolutions, Catalog. No. 1099-MAP2); anti-CDK5 mouse monoclonal IgG antibody (clone 4E4; Novus Bio, Catalog No. NBP2-37602); anti-GSK3β mouse monoclonal IgG antibody (clone D5C5Z; Novus Bio, catalog No. NBP147470S); anti-PS1 C-terminal (CTF) rabbit monoclonal IgG antibody (clone EP2000Y; Abcam, Catalog No. ab76083); anti-PS1 N-terminal (NTF) rabbit polyclonal IgG antibody (231-f; made in the Yankner lab); anti-Nicastrin mouse monoclonal IgG antibody (clone 9C3; Biolegend, Catalog No. 852301); anti-Nicastrin rabbit polyclonal IgG antibody (Sigma Millipore, Catalog No. N1660); anti-PEN2 rabbit polyclonal IgG antibody (ProScience, Catalog No.3981); anti-PEN2 rabbit monoclonal IgG antibody (clone EPR9200; Abcam, Catalog No. ab154830); antiPEN2 rabbit polyclonal IgG (ProScience, Catalog No. 3981); antiTransferrin receptor mouse monoclonal IgG antibody (clone H68.4; ThermoFisher Scientific, Catalog No. 13-6800); anti-BiP/GRP78 mouse monoclonal IgG (clone C38; ThermoFisher Scientific, clone C38, CatalogNo.

Techniques: Immunolabeling, Activation Assay, Marker, Expressing

Journal: Nature communications

Article Title: A neurodegeneration checkpoint mediated by REST protects against the onset of Alzheimer's disease.

doi: 10.1038/s41467-023-42704-6

Figure Lengend Snippet: Fig. 5 | REST suppresses the tau kinases CDK5 and GSK3β. a, b Loss of REST in excitatory neurons increases CDK5 expression in cortex and hippocampus. aImmunolabelingforCDK5(green)andthe neuronalmarkerMAP2(magenta)in CA1 neurons of the hippocampus in 9-month-old 3xTg and 3xTg;cKO mice. b Quantification of CDK5 immunofluorescence intensity in the hippocampus and cortex of 9-month-old 3xTg (n = 4) and 3xTg;cKO (n = 4) mice. c, d Loss of a single REST allele increases CDK5 expression. c Immunolabeling for CDK5 (green) and MAP2 (magenta) in 29-month-old 3xTg and 3xTg;GT (heterozygous REST null) mice. d Quantification of CDK5 immunofluorescence intensity in 28–29-month-old 3xTg (n = 6) and 3xTg;GT (n = 6) mice. e, f Loss of REST in excitatory neurons increases GSKβ expression in cortex and hippocampus. e Immunolabeling for GSK3β (green) and MAP2 (magenta) in hippocampal CA1 neurons in 9-month-old 3xTg and 3xTg;cKO mice. f Quantification of GSK3β immunofluorescenceintensityin9-month-old3xTg(n = 4)and3xTg;cKO (n = 4) mice. g, h Loss of a single REST allele increases GSK3β expression.

Article Snippet: Additional primary antibodies were as follows: anti-human Aβ rabbit monoclonal IgG antibody (Cell Signaling, Cat. No. 8243); antihuman APP mouse monoclonal IgG antibody (clone 6E10; Covance, Catalog No. SIG-39320); anti-actin mouse monoclonal IgG antibody (clone ACTN05 (C4); ThermoFisher Scientific, CatalogNo.MA5-11869); anti-NeuN mouse monoclonal IgG antibody (clone A60, Millipore, MAB377); anti-MAP2 goat polyclonal IgG antibody (PhosphoSolutions, Catalog. No. 1099-MAP2); anti-CDK5 mouse monoclonal IgG antibody (clone 4E4; Novus Bio, Catalog No. NBP2-37602); anti-GSK3β mouse monoclonal IgG antibody (clone D5C5Z; Novus Bio, catalog No. NBP147470S); anti-PS1 C-terminal (CTF) rabbit monoclonal IgG antibody (clone EP2000Y; Abcam, Catalog No. ab76083); anti-PS1 N-terminal (NTF) rabbit polyclonal IgG antibody (231-f; made in the Yankner lab); anti-Nicastrin mouse monoclonal IgG antibody (clone 9C3; Biolegend, Catalog No. 852301); anti-Nicastrin rabbit polyclonal IgG antibody (Sigma Millipore, Catalog No. N1660); anti-PEN2 rabbit polyclonal IgG antibody (ProScience, Catalog No.3981); anti-PEN2 rabbit monoclonal IgG antibody (clone EPR9200; Abcam, Catalog No. ab154830); antiPEN2 rabbit polyclonal IgG (ProScience, Catalog No. 3981); antiTransferrin receptor mouse monoclonal IgG antibody (clone H68.4; ThermoFisher Scientific, Catalog No. 13-6800); anti-BiP/GRP78 mouse monoclonal IgG (clone C38; ThermoFisher Scientific, clone C38, CatalogNo.

Techniques: Expressing, Immunolabeling

Journal: Journal of Dental Sciences

Article Title: Overexpression of sprouty 1 protein in human oral squamous cell carcinogenesis

doi: 10.1016/j.jds.2020.07.013

Figure Lengend Snippet: Western blot analyses: phosphor-ERK and total-ERK in human oral cancer cell lines as compared with a primary culture of normal oral mucosa (HOK). Upregulation of phosphor-ERK (normalized to total-ERK) in OECM1 and Ca922 cell lines as compared with HOK, and a slightly decreased expression in SAS as compared with HOK. Results were quantified using densitometric analysis, normalized to the level of β-actin, and expressed as a fold change relative to the normal oral mucosa. Bars represent means ± standard deviation of the mean (∗ P < 0.05). A representative result of three independent experiments is shown.

Article Snippet: Further, samples were analyzed using 10% SDS-PAGE (Sigma-Aldrich) gels, and the proteins were transmitted onto a PVDF membrane (Sigma-Aldrich) using Bio-Rad's transblot with primary antibodies against phosphor-ERK (Boster Biological Technology, CA, USA; Cat. No. P00104; 1:1000) and total-ERK (Boster Biological Technology; Cat. No. P00104; 1:1000), with species specificity for human tissues and an observed molecular weight of 42–44 kDa; and β-actin (Sigma-Aldrich; 1:1000), followed by horseradish peroxidase (HRP)-conjugated secondary antibodies (Sigma-Aldrich; 1:5000).

Techniques: Western Blot, Expressing, Standard Deviation

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