Fig. 4

circTmcc1 regulates EAAT2 transcriptions via the p65-NF-κB transcriptional activator complex. A Prediction of a protein factor that can regulate the transcription of the gene affected by circTmcc1 downregulation. The color bars exhibited the BART evaluation using the Irwin–Hall p-value and the ChEA3 evaluation using the integrated Score Rank, respectively. B The possibility of interaction between circTmcc1 and the transcription factor was predicted by STRING. C The interaction between circTmcc1 and p65 NF-κB, CREB, and c-Jun were confirmed through RNA–protein interaction in C8-D1a cells. D 293T cells were transfected with the mouse EAAT2 (Slc1a2) promoter luciferase construct, along with expression vectors for p65 and CREB following circTmcc1 knockdown. The changes of EAAT2 (Slc1a2) promoter activity were measured and reported as the mean ± SEM (n = 3). E Confirmation of changes in the expression of EAAT2 protein, a glutamate transporter in C8-D1a cells treated with circTmcc1 siRNAs, are described as the mean ± SEM (n = 3). F Immunocytochemical images of p-NF-κB/EAAT2 expression in circTmcc1-downregulated C8-D1a cells. The representative cells from three independent cultures (n = 3). G Immunocytochemical images of CREB/EAAT2 expression in circTmcc1-downregulated C8-D1a cells. The representative cells from three independent cultures (n = 3). An unpaired two-tailed t-test with Welch’s correction was used for statistical analysis. ns, not significant, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001