Fig. 4: Regulation of Bdnf transcription by circTshz2-2 and the YY1 transcriptional repressor complex.

A Illustration describing the mouse Bdnf gene and transcript variants, and the changes in the levels of Bdnf mRNA isoforms following circTshz2-2 knockdown in differentiated (Diff.) Neuro-2A cells at day 5. The alternative splicing sites in the mouse Bdnf gene are shown by the black bar. The data are reported as the mean ± SEM (n = 3). siCtr indicates the negative control siRNA and statistical significance was analyzed using an unpaired two-tailed t-test with Welch’s correction; ns not significant, *p < 0.05, **p < 0.01, ***p < 0.001. B The prediction of possible protein factors which may regulate the transcription of those genes affected by circTshz2-2-depletion in the RNA-seq data. The color bar represents the Irwin-Hall p value for the BART evaluation and the Integrated Score Rank from ChEA3, respectively. C The interaction probability between circTshz2-2 and specific transcriptional regulators was predicted using RPIseq. The color bar represents the interaction probability score (0–1). RF and SVM indicate random forest and support vector forest, respectively. D The interaction probability between circTshz2-2 and TSHZ2 co-factors or RNA-binding proteins was predicted by RPIseq. The color bar indicates the interaction probability score (0–1). E Confirmation of the RNA-protein interactions between circTshz2-2 and YY1 (upper) or CTBP (lower) in differentiated Neuro-2A cells at day 5. The data are from a representative experiment (n = 3). The band marked with an asterisk is circTshz2-1. IP indicates immunoprecipitation. F Illustration of the promoter-like signature (PLS) in the sequences of the mouse Bdnf gene from exons 4 through 6. The PLS data were obtained from the Candidate Cis-regulatory Elements database in ENCODE. The YY1 binding core motif and polycomb group (PcG) response elements are highlighted in red and green, respectively.