Fig. 3

circTmcc1 regulates intracellular molecular mechanisms in hyperammonemia-exposed astrocytes. A Volcano plot for gene with altered expression changes that differed by knockdown of circTmcc1 in C8-D1a cells exposed to ammonia. Those genes with p < 0.05 and with > 50% expression changes are indicated by colored dots. siCtr indicates control siRNA whereas sicircTmcc1 indicates circTmcc1 siRNA. B Gene expression following the circTmcc1 knockdown in C8-D1a cells. C The gene expression changes related to GPCR and cellular metabolism following circTmcc1 knockdown. In B and C, the heat map represents the difference in the expression compared to that in the negative control siRNA-treated C8-D1a cells. D The Gene Ontology (GO) analysis of differentially expressed 287 protein-coding genes in volcano plot. The top 10 GO terms of biological process appear in FDR q-value. E The changes in cAMP-related mRNA (Fzd6 and Adcy1) levels following circTmcc1 knockdown in C8-D1a cells were measured and represented as the mean ± SEM (n = 3). F Measurement of cAMP concentration in C8-D1a cells after circTmcc1 knockdown. The data are expressed as three independent experiments in duplicate. G The measurement of FZD6, ADCY1, p-PKA and p-CREB protein levels following circTmcc1 knockdown in C8-D1a cells were measured and are depicted as the mean ± SEM (n = 3). p-PKA and p-CREB protein levels compared with total PKA and CREB protein levels. An unpaired two-tailed t-test with Welch’s correction was used for statistical analysis. ns, not significant, *p < 0.05, **p < 0.01