Figure 1.

NHD minigene shows exon 3 skipping. (A) Schematic diagram of the TREM2 exon 2–4 minigene. The minigene is inserted downstream of the EGFP cDNA. The positions of the primer set used to detect splicing are indicated by arrows. A red arrowhead indicates the position of the 5′ splice site mutation (c.482 + 2 T > C) found in NHD. (B) Splicing assay of WT and NHD minigenes transfected into HEK cells. Splicing patterns were detected by RT-PCR using the primer set indicated in (A). (C) Unmodified U1 and U7 snRNAs did not alter the splicing pattern of the NHD minigene. The NHD minigene was transfected with either an empty vector, unmodified U1 snRNA, or unmodified U7 snRNA. Splicing patters were detected as indicated in (B). Exon 3 inclusion was not induced by co-expression of U1 or U7 snRNA. Original gel images are shown in Supplementary Fig. S8.