Fig. 2: CircEZH2 encodes the peptide EZH2-92aa.

a Illustration of the circEZH2-encoded peptide EZH2-92aa. The unique C-terminus of EZH2-92aa (in red) is produced by an ORF spanning more than 360° in circEZH2. A custom antibody against the indicated unique C-terminal sequence was produced. b 293T cells transfected with the circEZH2 plasmid were subjected to polysome profiling. CircEZH2 was detected by qPCR in the indicated fractions. CircHIPK3 and EZH2 served as the negative and positive controls, respectively. c Illustration of endogenous circEZH2, the del-ATG circEZH2 construct and the linearized EZH2-92aa-3×Flag construct. d Immunoblot of cells overexpressing the above constructs using the custom anti-EZH2-92aa and anti-flag antibodies. VA, vector alone. e Identification of the unique C-terminal peptide sequence of EZH2-92aa in 293T cells overexpressing circEZH2 and in MES28 GSCs by MS. f Top, EZH2-92aa expression levels were measured in NHAs and several established GSC lines. Bottom, immunoblot of EZH2-92aa in seven randomly selected paired GBM samples using the custom anti-EZH2-92aa antibody. g Semiquantitative analysis of the EZH2-92aa expression level based on greyscale analysis in the aforementioned cohort of 63 high-grade glioma samples. Two-sided paired t test, P = 3.14e−05. h Survival analysis of patients stratified by EZH2-92aa expression (with the median expression score as the cut-off value) in the same cohort. Log-rank test, P = 0.0066. The data in (b), (d), (e) and (f) are pooled from three independent experiments and are presented as the mean ± SD. Source data are provided as a Source data file.