Fig. 2

Identification of circPTPN4. (A) Schematic image of circPTPN4 derived from PTPN4. (B) Verification of circPTPN4 by amplifying with divergent primers. (C) Sanger sequencing confirmed the back-splicing junction sequence of circPTPN4. (D) Relative circPTPN4 and β-actin expression after treatment with RNase R. (E and F) Relative circPTPN4 expression in pectoralis major (PEM) and soleus (SOL) of 7-week-old Xinghua chicken detected by RNA-seq (E) and qPCR (F). (G) Tissue expression profiles of circPTPN4. The horizontal axis and vertical axis indicate different tissues and their relative expression values, respectively. (H) The distribution of circPTPN4 in the cytoplasm and nuclei of chicken primary myoblast (CPM) was determined by qRT-PCR. GAPDH and U6 serve as cytoplasmic and nuclear localization controls, respectively. (I) The transcriptional activity of the PTPN4 core promoter region. (J) Chromatin immunoprecipitation (ChIP) analysis of the binding capacity of FOXA2 to the PTPN4 core promoter region. (K) Relative FOXA2, circPTPN4 and circPTPN4 expression with FOXA2 overexpression. Results are presented as mean ± SEM. In panels (D to H, and K), the statistical significance of differences between means was assessed using paired t-tests. In panels (I and J), ANOVA followed by Dunnett’s test was used. (*P < 0.05; **P < 0.01)