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: The Journal of Neuroscience

Article Title: β-Amyloid-Stimulated Microglia Induce Neuron Death via Synergistic Stimulation of Tumor Necrosis Factor α and NMDA Receptors

doi: 10.1523/JNEUROSCI.4998-04.2005

Figure Lengend Snippet: Conditioned media from Aβ-stimulated microglia induce neuron death in a TNFα-dependent manner. Conditioned Neurobasal media (CM) from primary mouse microglia (400 cells/mm2) stimulated for 48 h with or without (control) immobilized Aβ1-42 fibrils (48 pmol/mm2) was generated. A, Control and Aβ-stimulated conditioned media were applied to mouse cortical neuron cultures (E16; 7 d in vitro) for 72 h. Neurons were fixed in 4% paraformaldehyde, stained using anti-MAP2 antibody, and counted. Neurons from four fields/conditions were counted in quadruplicate wells and averaged ± SEM. The graph is representative of three independent experiments. B, Conditioned media were collected, and TNFα concentrations were determined via commercial ELISA. The graph is representative of three independent experiments. C, Cortical neurons were cultured for 72 h with unstimulated and Aβ-stimulated conditioned medium in the absence or presence of soluble TNFRI (0.1 μg/ml). Neurons were fixed, stained using anti-MAP2 antibody, counted as above, and averaged ± SEM. The graph is representative of three independent experiments. D, Media from plates containing increasing concentrations of immobilized Aβ (0, 48, 96, and 192 pmol Aβ1-42/mm2) alone were transferred to cortical neurons for 72 h. Neurons were fixed and counted, and statistical significance was determined as above. *p < 0.001 from control; **p < 0.001 from CM.

Article Snippet: Neurons were double labeled using anti-MAP2 antibody and anti-iNOS antibody (Alexis Biochemicals).

Techniques: Generated, In Vitro, Staining, Enzyme-linked Immunosorbent Assay, Cell Culture

Journal: The Journal of Neuroscience

Article Title: β-Amyloid-Stimulated Microglia Induce Neuron Death via Synergistic Stimulation of Tumor Necrosis Factor α and NMDA Receptors

doi: 10.1523/JNEUROSCI.4998-04.2005

Figure Lengend Snippet: Conditioned media-dependent death requires NMDA receptor activity. A, Conditioned Neurobasal media (CM) from primary mouse microglia (400 cells/mm2) stimulated for 48 h with or without (control) immobilized Aβ1-42 fibrils (48 pmol/mm2) were generated, and micromolar glutamate concentrations were calculated from the media. The graph is the average ± SEM of three independent experiments. Cortical neurons (E16; 7 d in vitro) were cultured for 72 h with conditioned medium from primary mouse microglia that were unstimulated or stimulated (CM) 48 h with Aβ1-42 fibrils in the absence or presence of 1 μm memantine and 10 μm APV (B) or 1, 5, and 50 μm NBQX (C). Neurons were then fixed, stained using anti-MAP2 antibody, and counted. Neurons from four fields/conditions were counted in quadruplicate wells and averaged ± SEM. Graphs are representative of three independent experiments. *p < 0.001 from control; **p < 0.001 from CM.

Article Snippet: Neurons were double labeled using anti-MAP2 antibody and anti-iNOS antibody (Alexis Biochemicals).

Techniques: Activity Assay, Generated, In Vitro, Cell Culture, Staining

Journal: The Journal of Neuroscience

Article Title: β-Amyloid-Stimulated Microglia Induce Neuron Death via Synergistic Stimulation of Tumor Necrosis Factor α and NMDA Receptors

doi: 10.1523/JNEUROSCI.4998-04.2005

Figure Lengend Snippet: TNFα and glutamate/NMDA synergistically stimulate neuron death. Mouse cortical neurons (E16; 7 d in vitro) were cultured in the absence or presence of NMDA (50, 100 μm) (A), glutamate (25, 50, 100 μm) (B), and mouse TNFα (5, 50 ng/ml). Stimuli were added to neurons for 72 h and then cells were fixed, stained using anti-MAP2 antibody, and counted. Neurons from four fields/conditions were counted in quadruplicate wells and averaged ± SEM. Graphs are representative of three independent experiments. *p < 0.05 from control; **p < 0.001 from control.

Article Snippet: Neurons were double labeled using anti-MAP2 antibody and anti-iNOS antibody (Alexis Biochemicals).

Techniques: In Vitro, Cell Culture, Staining

Journal: The Journal of Neuroscience

Article Title: β-Amyloid-Stimulated Microglia Induce Neuron Death via Synergistic Stimulation of Tumor Necrosis Factor α and NMDA Receptors

doi: 10.1523/JNEUROSCI.4998-04.2005

Figure Lengend Snippet: TNFα plus NMDA induces increased neuronal iNOS immunoreactivity. Neurons were unstimulated (A-D) or stimulated (E-H) 72 h with TNFα (50 ng/ml) plus 100 μm NMDA and then fixed in 4% paraformaldehyde. Cultures were double labeled using anti-MAP2 and anti-iNOS antibodies with FITC and Texas Red-conjugated secondary antibodies, respectively. Cultures were mounted in DAPI containing mounting media for confocal imaging. A, E, Anti-MAP2; B, F, anti-iNOS; C, G, DAPI; D, merge of A-C; H, merge of E-G. Images are representative of three independent experiments.

Article Snippet: Neurons were double labeled using anti-MAP2 antibody and anti-iNOS antibody (Alexis Biochemicals).

Techniques: Labeling, Imaging