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. 2022 Jul 2;70(11):2079–2092. doi: 10.1002/glia.24239

FIGURE 5.

FIGURE 5

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)