mouse lncrna microarray v2.0 (8 60k)  (Arraystar inc)

Journal: Glia

Article Title: The LncRNA AK018453 regulates TRAP1/Smad signaling in IL‐17‐activated astrocytes: A potential role in EAE pathogenesis

doi: 10.1002/glia.24239

Figure Lengend Snippet: LncRNA AK018453 is upregulated and regulates TRPA1 expression in activated astrocytes by IL‐17. (a) Comparative lncRNA array analysis indicated that differentially upregulated lncRNAs occurred in the primary astrocytes treated with IL‐17 (50 ng/ml) at 3 and 6 h (2.0‐fold higher than DMEM group). (b) Hot map showed some differentially‐expressed lncRNAs in astrocytes stimulated with IL‐17. Red box circled differentially co‐upregulated lncRNA AK018453. (c) Real‐time PCR assay was employed to verify lncRNA AK018453 expression in primary mouse astrocytes treated by IL‐17 and spinal cords from EAE mice. Results were represented as mean ± SEM. ** p < .01 and *** p < .001 versus 0 h group. # p < .05 and ## p < .01 versus NC group ( n = 3). (d and e) RIP assay was performed to investigate the interaction of AK018453 with NF‐κB p65 and CBP/P300 in astrocytes stimulated with IL‐17 for 6 h, or prior to infection by lentiviruses AK018453‐shRNA (AK‐shRNA) and ctrl‐shRNA for 72 h. The data were from three independent experiments. ** p < .01 and *** p < .001 versus DMEM. # p < .05 versus ctrl‐shRNA + IL‐17 group ( n = 3). (f and g) ChIP assay analysis of NF‐κB p65, CBP/P300, H3K27ac, and RNA pol II enrichment on the promoter of TRAP1 gene in astrocytes stimulation with IL‐17 for 6 h, or prior to infection by lentiviruses AK‐shRNA and ctrl‐shRNA for 72 h. The data were from three independent experiments and represented as mean ± SEM. * p < .05, ** p < .01, *** p < .001 versus DMEM; # p < .05 and ## p < .01 versus ctrl‐shRNA + IL‐17 group ( n = 3)

Article Snippet: In brief, Mouse LncRNA Microarray v2.0 (8 × 60K, Arraystar) was performed by Kangchen Bio‐tech (Shanghai, China) to measure the global profiling of mouse LncRNAs in primary mouse astrocytes treated with or without IL‐17.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Infection, shRNA

Journal: Glia

Article Title: The LncRNA AK018453 regulates TRAP1/Smad signaling in IL‐17‐activated astrocytes: A potential role in EAE pathogenesis

doi: 10.1002/glia.24239

Figure Lengend Snippet: AK018453 knockdown lessens the pathology of EAE mice. On day 7 after administration with recombinant lentivirus carrying GFAP promoter and AK‐shRNA or ctrl‐shRNA sequence, respectively, mice were immunized with or without MOG 35–55 for 21 days. (a) The level of lncRNA AK018453 was determined by real‐time PCR assay. *** p < .001 versus NC group; # p < .05 versus ctrl‐shRNA group ( n = 10 mice/group). (b) The clinical scores for EAE mice infected with AK‐shRNA or ctrl‐shRNA ( n = 10 mice per group). *** p < .001 versus NC group; ## p < .01 versus ctrl‐shRNA group. (c) Western blot assay was used to determine the protein levels of TRAP1, Smad4, p‐Smad2/3, Smad2/3, and GFAP in the spinal cords. (d and e) The production of TNF‐α, CXCL10, and MCP‐1 in the spinal cords and peripheral blood from the AK‐shRNA and ctrl‐shRNA mice was measured by real‐time PCR assay and ELISA assay, respectively. The data are represented as mean ± SEM. *** p < .001 versus NC group; # p < .05, ## p < .01, and ### p < .001 versus ctrl‐shRNA group ( n = 10 mice/group). (f) Hematoxylin and eosin (H&E) staining was designed to observe the infiltrations of inflammatory cells in spinal cords (scale bars, 50 μm). (g) Luxol fast blue (LFB) staining was utilized to test the medullary sheath damages from spinal cords (scale bars, 50 μm). Red box areas in the upper rows are presented enlarged underneath. (h) Electron microscope (EM) was employed to investigate the severe disruption or mild loosening of the medullary sheath in spinal cords. Red arrow shows the changes of the medullary sheath. Scale bars, 1 μm

Article Snippet: In brief, Mouse LncRNA Microarray v2.0 (8 × 60K, Arraystar) was performed by Kangchen Bio‐tech (Shanghai, China) to measure the global profiling of mouse LncRNAs in primary mouse astrocytes treated with or without IL‐17.

Techniques: Knockdown, Recombinant, shRNA, Sequencing, Real-time Polymerase Chain Reaction, Infection, Western Blot, Enzyme-linked Immunosorbent Assay, Staining, Microscopy, Disruption

Journal: Glia

Article Title: The LncRNA AK018453 regulates TRAP1/Smad signaling in IL‐17‐activated astrocytes: A potential role in EAE pathogenesis

doi: 10.1002/glia.24239

Figure Lengend Snippet: Schematic representation of lncRNA AK018453 contributing to the pathology of EAE mice via regulating TRAP1/Smad pathway. Response to IL‐17 stimulation, AK018453 is upregulated in astrocytes, which regulates epigenetically the expression of TRAP1 through interacting with CBP/P300, in turn promotes the production of pro‐inflammatory cytokines such as TNF‐α, CXCL10, and MCP‐1, thus aggravating the EAE progression

Article Snippet: In brief, Mouse LncRNA Microarray v2.0 (8 × 60K, Arraystar) was performed by Kangchen Bio‐tech (Shanghai, China) to measure the global profiling of mouse LncRNAs in primary mouse astrocytes treated with or without IL‐17.

Techniques: Expressing