Extended Data Fig. 2: Characterization of dual-luciferase EXSISERSMAPT:10NLuc-11Fluc.
From: Non-invasive and high-throughput interrogation of exon-specific isoform expression
a, EXSISERSMAPT:10NLuc/11FLuc, 0N4R CDS containing the constitutive intron 11-12 was cloned into a mouse Pgk1 promoter-driven plasmid including the intron between exon 11 and 12. Additionally, mutations that disrupt intein splicing (C-terminal Asn→Ala mutation) were created to show their importance for efficient protein splicing. b, Immunoblots against pan-tau from cells expressing the two constructs described in a with CCs, either with an active or a catalytically inactive intein. Data shown represent 2 independent experiments. c, Increasing amounts of Pgk1 promoter-driven EXSISERSMAPT:10NLuc/11FLuc 0N4R CDS, driving the expression of both luciferases at 1:1 stoichiometry, were transfected into HEK293T cells. The resulting NLuc signal (RLU) is plotted against the FLuc signal (RLU) with a linear fit through the data points, revealing that inclusion of both exons results in NLuc signals that are ~30-fold (95% confidence interval (CI) from 28.08 to 30.63) stronger than FLuc. The reference curve was obtained from one experiment. Unprocessed blots are provided in Source Data Extended Data Fig. 2.
