tgm2 inhibitor cystamine  (MedChemExpress)

Journal: CNS Neuroscience & Therapeutics

Article Title: Inhibition of Transglutaminase 2 Preserves Blood–Brain Barrier Integrity and Improves Neurological Outcomes After Experimental Traumatic Brain Injury in Mice

doi: 10.1002/cns.70887

Figure Lengend Snippet: The expression of TGM2 and disruption of BBB integrity in Bend.3 after OGD. (A) Heatmap of Tgm2 and Tagln2 mRNA expression after TBI at 12 h, 24 h, and 72 h vs. Sham groups. (B, C) Protein quantification in four brain cell lines, n = 6 (independent cell culture preparations) in each group. (D) Tgm2 mRNA levels across four cell lines, n = 3 (independent cell culture preparations) in each group. (E–H) CLAUDIN‐5/ZO‐1 protein expression in OGD‐24 h vs. the control group. (I, J) Immunofluorescence staining of TGM2 after OGD at 24 h, Scale bar = 50 μm. (K, L) TGM2 protein quantification in OGD‐24 h vs. the control group, n = 6 (independent cell culture preparations) in each group. All data are shown as the mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. Control/OGD group. BV‐2: Mouse microglial cell line. Bend.3: Mouse brain microvascular endothelial cell line. C8: Mouse cerebellar astrocyte cell line. HT22: Mouse hippocampal neuronal cell lines. OGD: Oxygen Glucose Deprivation. TBI, Traumatic brain injury.

Article Snippet: The TGM2 inhibitor Cystamine (purchased from MedChemExpress, MCE) was dissolved in phosphate‐buffered saline (PBS) at a concentration of 100 mg/mL according to the manufacturer's instructions.

Techniques: Expressing, Disruption, Cell Culture, Control, Immunofluorescence, Staining

Journal: CNS Neuroscience & Therapeutics

Article Title: Inhibition of Transglutaminase 2 Preserves Blood–Brain Barrier Integrity and Improves Neurological Outcomes After Experimental Traumatic Brain Injury in Mice

doi: 10.1002/cns.70887

Figure Lengend Snippet: The expression of TGM2 and disruption of BBB integrity after TBI. (A, B) Time course of TGM2 expression in the brain after TBI, n = 6 mice/group. (C, D) Neurological deficits assessed by mNSS and wire‐hanging tests, n = 6 mice/group. (E) Dual immunofluorescence of TGM2(red) and lectin(green) for co‐localization, Scale bar = 25 μm. (F–I) CLAUDIN‐5 and ZO‐1 protein quantification after TBI, n = 6 mice/group. (J) Brain water content quantification, n = 6 mice/group. (K) Immunofluorescence shows CLAUDIN‐5 and lectin co‐staining. Scale bar = 25 μm. All data are shown as the mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. Sham/TBI‐48 h group. TBI: Traumatic brain injury; mNSS: Modified neurologic severity score.

Article Snippet: The TGM2 inhibitor Cystamine (purchased from MedChemExpress, MCE) was dissolved in phosphate‐buffered saline (PBS) at a concentration of 100 mg/mL according to the manufacturer's instructions.

Techniques: Expressing, Disruption, Immunofluorescence, Staining, Modification

Journal: CNS Neuroscience & Therapeutics

Article Title: Inhibition of Transglutaminase 2 Preserves Blood–Brain Barrier Integrity and Improves Neurological Outcomes After Experimental Traumatic Brain Injury in Mice

doi: 10.1002/cns.70887

Figure Lengend Snippet: TGM2‐KD can regulate BBB relative protein expression in ECs. (A, C) TGM2 knocked down efficiency in the Bend.3, n = 6 (independent cell culture preparations) in each group. (B, D) OGD‐induced TGM2 protein alterations in shCtrl vs. shTGM2, n = 6 (independent cell culture preparations) in each group. (E–H) mRNA expression levels of Tgm2 / Occludin / Claudin‐5 / Zo‐1 , n = 3 (independent cell culture preparations) in each group. (I–L) CLAUDIN‐5/ZO‐1 protein rescued by shTGM2 post‐OGD, n = 6 (independent cell culture preparations) in each group. All data are shown as the mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. shCtrl/shTGM2 OGD24h +/− group. KD: Knockdown; BBB: Blood–brain barrier; ECs: Endothelial cells; OGD: Oxygen Glucose Deprivation.

Article Snippet: The TGM2 inhibitor Cystamine (purchased from MedChemExpress, MCE) was dissolved in phosphate‐buffered saline (PBS) at a concentration of 100 mg/mL according to the manufacturer's instructions.

Techniques: Expressing, Cell Culture, Knockdown

Journal: CNS Neuroscience & Therapeutics

Article Title: Inhibition of Transglutaminase 2 Preserves Blood–Brain Barrier Integrity and Improves Neurological Outcomes After Experimental Traumatic Brain Injury in Mice

doi: 10.1002/cns.70887

Figure Lengend Snippet: TGM2‐KD down‐regulates MMP9 expression through the IL‐17 pathway in Bend.3 after OGD. (A) Volcano plot of DEGs in TGM2‐KD vs. NC‐OGD group, with red points representing up‐regulated genes and blue points representing down‐regulated genes. (B, C) GO and KEGG enrichment analysis of down‐regulated DEGs. (D) The top 10 hub genes calculated by cytoHubba. (E, F) MMP9 protein suppression by shTGM2 post‐OGD, n = 6 (independent cell culture preparations) in each group. (G) mRNA expression level of Mmp9 in shTGM2 after OGD, n = 3 (independent cell culture preparations) in each group. All data are shown as the mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. shCtrl+shTGM2 OGD24h +/− group. KD: Knockdown; OGD: Oxygen Glucose Deprivation; DEG: Differentially expressed genes. GO: Gene Ontology; KEGG: Kyoto Encyclopedia of Genes and Genomes; PPI: Protein–Protein Interaction.

Article Snippet: The TGM2 inhibitor Cystamine (purchased from MedChemExpress, MCE) was dissolved in phosphate‐buffered saline (PBS) at a concentration of 100 mg/mL according to the manufacturer's instructions.

Techniques: Expressing, Cell Culture, Knockdown

Journal: CNS Neuroscience & Therapeutics

Article Title: Inhibition of Transglutaminase 2 Preserves Blood–Brain Barrier Integrity and Improves Neurological Outcomes After Experimental Traumatic Brain Injury in Mice

doi: 10.1002/cns.70887

Figure Lengend Snippet: Effect of Cystamine (TGM2 inhibitor) on the BBB after TBI in mice. (A) Schematic of drug administration in mice. (B–F) mRNA expression of AQP4 / Occludin / Claudin‐5 / Zo‐1 / Mmp9 , n = 3 mice/group. (G–I) Representative Western blot imaging of AQP4、CLAUDIN‐5, and ZO‐1. (K–L) Protein quantification shows cystamine‐mediated AQP4 suppression and CLAUDIN‐5/ZO‐1 restoration, n = 6 mice/group. (M) Quantification of Evans blue extravasation, n = 6 mice/group. (N, O) Evans blue extravasation imaging, Scale bar = 20 μm. All data are shown as the mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. Sham and vehicle/cystamine group. BBB: Blood–brain barrier; TBI: Traumatic brain injury.

Article Snippet: The TGM2 inhibitor Cystamine (purchased from MedChemExpress, MCE) was dissolved in phosphate‐buffered saline (PBS) at a concentration of 100 mg/mL according to the manufacturer's instructions.

Techniques: Expressing, Western Blot, Imaging

Journal: CNS Neuroscience & Therapeutics

Article Title: Inhibition of Transglutaminase 2 Preserves Blood–Brain Barrier Integrity and Improves Neurological Outcomes After Experimental Traumatic Brain Injury in Mice

doi: 10.1002/cns.70887

Figure Lengend Snippet: Effect of Cystamine (TGM2 inhibitor) on neuromotor function after TBI in mice. (A–C) Rotarod latency, n = 6 mice/group. (D) mNSS score, n = 6 mice/group. (E, F) Open field test (Trajectory diagram; trajectory heatmap; total distance), n = 6 mice/group. (G) Morris water maze trajectory diagram. (H–K) Morris water maze statistics of platform crossings, the time spent in the target quadrant, distance in the target quadrant, and swim speed, n = 6 mice/group. (L) Beam‐walk scores, n = 6 mice/group. All data are shown as the mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. Sham and vehicle/cystamine group. mNSS: Modified neurologic severity score.

Article Snippet: The TGM2 inhibitor Cystamine (purchased from MedChemExpress, MCE) was dissolved in phosphate‐buffered saline (PBS) at a concentration of 100 mg/mL according to the manufacturer's instructions.

Techniques: Modification