Fig. 4

Identification of the splice site variant using the minigene splicing assay. A A schematic of the pSPL3 vector with cloned wild type and mutant type GSDME, including partial intron 6, exon 7 (yellow), intron 7, exon 8 (green), intron 8, exon 9 (red), and partial intron 9. A schematic of the splice products with the wild type splicing profile (bottom left) and mutant type splice variant profile (bottom right). B Agarose gel electrophoresis of empty pSPL3 vector (263 bp), c.1183 + 1 G > C variant (465 bp), and wild type (658 bp). C Sanger sequencing chromatograms of the minigene splicing assay products. Wild type (top); mutant type with skipping of exon 8 (bottom)