Fig. 5

Minigene splicing assay for GAS2. A Structure of GAS2 gene and the surrounding area of rs11026723. Exons are depicted by the thick lines. This figure was obtained from the UCSC genome browser. B Isoforms of GAS2. Isoform structures are depicted using GSDS 2.0 [23]. C Boxplots displaying gene expressions in the 67 Japanese B cell lines. The expression levels of the entire GAS2 gene (left) and isoforms are presented by the genotype (GG (n = 16), GA (n = 36), and AA (n = 15)). D Agarose gel electrophoresis of RT-PCR products expressed from GAS2 (exon 1) minigenes in HEK293 cells. In this experiment, a modified pSPL3 plasmid was utilized (Additional file 1: Fig. S2). M: marker (DNA ladder One (Nacalai)). Lane 1: pSPL3 + GAS2 (IVS1 + 3 A) (RT +); Lane 2: pSPL3 + GAS2 (IVS1 + 3G) (RT +); Lane 3: pSPL3 + GAS2 (IVS1 + 3 A) (RT −); Lane 4: pSPL3 + GAS2 (IVS1 + 3G) (RT −); Lane 5: pSPL3-GAS2 plasmid. A purified plasmid was used as the PCR template. Lane 6: Negative control. RT: Reverse transcription. The size of the upper band in lanes 1 and 2 was the same as the amplicon in lane 5, suggesting the presence of an unspliced transcript. Blue box: exon of pSPL3 plasmid; green box: exon 1 of GAS2