Fig. 1: Regulation of neuronal structures by circTshz2-2.

A Genomic information and conservation of circTshz2-2 in mouse Tshz2 locus (mm10). Genomic information and PhastCons data were obtained from the USCS Genome Browser. Back-splice junctions of circRNAs identified by Sanger sequencing are indicated as a silver triangle. B Changes in neural structures and neurite length following circTshz2-2 knockdown in differentiated (Diff.) Neuro-2A cells at day 5. siCtr denotes the negative control siRNA. C Changes in neural structures and total neurite length following circTshz2-2 knockdown in mouse primary cortical neurons (PCN) after 7 days in vitro (DIV 7). D Changes in the number of neurites from each soma following circTshz2-2 knockdown in PCN at DIV 7. The whiskers in graphs B–D represent the minimum and maximum values while the box line represents the first quartile, median, and third quartile, respectively. The “+“ indicates the mean. E Changes in neural complexity following circTshz2-2 knockdown in PCN at DIV 7. The number of intersections was analyzed using the Sholl analysis method as described in Fig. S9E and results are described as the mean (solid line) ± SEM (dot line). F Changes in SYP and PSD-95 protein levels following circTshz2-2 knockdown in PCN at DIV 7. Data are represented as the mean ± SEM (n = 3). Structural data for B–E represents the findings from at least three independent experiments (n = 3) with more than ten neurons analyzed per replicate. Statistical significance for B–F was determined using an unpaired two-tailed t-test with Welch’s correction. The data in E were evaluated using an ordinary two-way ANOVA test. *p < 0.05, **p < 0.01, ***p < 0.001.