Fig. 4: SRSF1 regulates alternative splicing of SRA1 exon 3 by binding to “GGAACAGGCATTGGAAGA” sequence in exon 3 to promote exon inclusion. | Cell Death Discovery

Fig. 4: SRSF1 regulates alternative splicing of SRA1 exon 3 by binding to “GGAACAGGCATTGGAAGA” sequence in exon 3 to promote exon inclusion.

From: SRSF1 promotes the inclusion of exon 3 of SRA1 and the invasion of hepatocellular carcinoma cells by interacting with exon 3 of SRA1pre-mRNA

Fig. 4

A RT-qPCR was used to detect the expression levels of different splicing factors in LO2, HepG2, and HCCLM3 cells. B Detect the mRNA expression of SRSF1, SRSF8, SRSF11 and SRA1-L in HCCs. C Correlation analysis of the expression levels of SRSF1, SRSF8, SRSF11 and SRA1-L. Use SPSS analysis tools for correlation analysis of gene expression. D Alternative splicing of exon 3 of SRA1 was examined in HCCLM3 with over-expression or knockdown of SRSF1, SRSF8, and SRSF11 by RT-PCR. E Graphical and specific primer design sites for pcDNA3.1-SRA1-minigene. The primers for the internal reference of exogenous SRA1-minigene were CMV-FP (FP), exon 2-RP (E2-RP), exon 3-RP (E3-RP) and exon 2/exon 3 splice junction-RP (E2-E4-RP), and the primers for RT-qPCR detection of exogenous SRA1 isoforms in HCCLM3. F The primers used to detect the binding segment of SRSF1 and its positions in SRA1 pre-mRNA in the CLIP experiment. The CLIP assays were applied to detect the binding between SRSF1 and SRA1 pre-mRNA. G The RNA fragment sequences utilized in the RNA pulldown assay and its positions in the SRA1 pre-mRNA. RNA pulldown assays were used to detect the binding between SRSF1 and the biotinylated SRA1 RNA fragments. H Graphical representation of the MS2-GFP-IP system and its mode of operation. The effect of SRSF1 binding to SRA1 was investigated by the MS2-GFP-IP system. Data are presented as mean ± S.D. (N = 3). The “*, **, ***” indicate “P < 0.05, 0.01, 0.001” versus the control group, respectively.

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