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. Author manuscript; available in PMC: 2018 Dec 7.
Published in final edited form as: Mol Cell. 2017 Nov 22;68(5):940–954.e3. doi: 10.1016/j.molcel.2017.10.034

Figure 6. Readthrough transcription generates endogenous circular RNAs from the human PAIP2 gene.

Figure 6

(A) Readthrough transcription from MATR3 as shown by nascent RNA-seq in human PA1 cells (Zhang et al., 2016). The downstream PAIP2 gene generates a circular RNA from exons 2 and 3 (purple). See also Figure S5. (B) Immunoblot to confirm depletion of CPSF3 (top) or CPSF4 (bottom) by shRNAs in PA1 cells. scr, scrambled shRNA control. Actin was used as a loading control. (C) Schematic showing qRT-PCR primers used to quantify transcripts from the MATR3-PAIP2 locus. (D and E) qRT-PCR quantification of transcripts from the MATR3-PAIP2 intergenic region (D) or the PAIP2 gene (E) in PA1 cells depleted of CPSF3 or CPSF4. Data are shown as mean±SD from three independent experiments. * p<0.05, ** p<0.01 (F) PA1 cells were treated for 8 h with a phosphorothioate-modified antisense oligodeoxynucleotide (ASO) complementary to the MATR3-PAIP2 intergenic region. (G) qRT-PCR was then used to quantify transcripts from the MATR3-PAIP2 intergenic region or the PAIP2 gene. Locations of primers are shown in F. Data are shown as mean±SD from three independent experiments. * p<0.05, ** p<0.01