Fig. 2

PRDM1 activates circ_0036412 transcription to facilitate the proliferation and inhibit the cell cycle of HCC cells in vitro. A UCSC (http://genome-asia.ucsc.edu/index.html) was used to predict the transcription factors regulating ETFA. B The expression of circ_0036412 was detected by qRT-PCR in HuH-7 and Hep 3B cells after the transfection of si-CTCF-1/2, si-PRDM1-1/2 or si-POLR2A-1/2 (One-way ANOVA, Dunnett). C JASPAR (https://jaspar.genereg.net/) predicted the binding sites of PRDM1 and ETFA promoter sequences. The site was located on the 1505–1515 fragment, and the sequence is CTCTCTCTCTC. D Luciferase reporter assay detected the interaction between PRDM1 and ETFA promoter in 293 T cells (Two-way ANOVA, Tukey). E–F ChIP (Student’s test) and DNA pulldown assays detected the interaction between PRDM1 and ETFA promoter in HuH-7 and Hep 3B cells. G, H EdU and CCK-8 assays evaluated cell proliferation in HuH-7 and Hep 3B cells after the transfection of si-circ_0036412-1/2 (One-way ANOVA, Dunnett). (I) TUNEL assay assessed cell apoptosis in HuH-7 and Hep 3B cells after the transfection of si-circ_0036412-1/2 (One-way ANOVA, Dunnett). J Flow cytometry analysis testified cell cycle in HuH-7 and Hep 3B cells after the transfection of si-circ_0036412-1/2 (One-way ANOVA, Dunnett). K, L The mRNA levels of CDC2 and CCNB1 were detected by qRT-PCR in HuH-7 and Hep 3B cells after the transfection of si-circ_0036412-1/2 (One-way ANOVA, Dunnett). **P < 0.01