Fig. 4

Genetic analysis and MAPT splicing assay. a Sanger sequencing of MAPT intron 9/exon10 boundary, displaying a reference sequence (top) and two consistent electropherograms (reverse and forward directions). b Schematic diagram depicting the design of MAPT mini gene construct with or without disease-causing mutation/variant. c Reverse Transcription Polymerase Chain Reaction (RT-PCR) of MAPT minigene transcripts (3 independent experiments). Complementary DNAs were collected from HeLa cells transfected with each plasmid. d Western blotting analysis of transfected HeLa cells in duplicate probed with anti-HA tag antibody (two independent experiments). Bands corresponding to 4 repeat tau (Exon 10 (+)) and 3 repeat tau (Exon 10 (−)) were observed. Anti-β-actin antibody was used as protein loading controls. RT(−): no reverse transcription. Mock indicates backbone plasmid vector lacking MAPT-related sequence. Exon 10 (+) and (−) indicate bands corresponding to exon 10 inclusion and exclusion