Fig. 1

Characterization of circAF4 in leukemia cells. a Schematic of the genomic loci of four circRNAs in AF4 gene. Arrows represent the length of indicated circRNAs. b Divergent primers were used for circRNA validation. c RNA was isolated from human leukemia cell line, RS4;11. The expressions of four circAF4 isoforms were validated by qTR-PCR. Expression was calculated using the 2^−ΔΔCT method and normalized to GAPDH. d The qTR-PCR product of circAF4 was validated by sanger sequencing. The arrow represents the unique junction site of circAF4. e qRT-PCR for the abundance of circAF4 and GAPDH mRNA in cells treated with RNase R. The amount of circAF4 and GAPDH mRNA were normalized to the value measured in the mock treatment. f Transcription was blocked by adding 2 μg ml⁻¹ actinomycin D to the RS4;11 cell culture medium. RNAs were extracted in the indicated time points. The qTR-PCR analysis was performed as described above. NS, not significant. *P < 0.05, ***P < 0.001. g The qTR-PCR analysis of the relative expression of circAF4 in leukemia patients and healthy people. NS, not significant. *P < 0.05, ***P < 0.001