Fig. 2: Live imaging and longitudinal readout of MAPT isoform expression.
From: Non-invasive and high-throughput interrogation of exon-specific isoform expression
a, EXSISERSMAPT:10NLuc-11FLuc HEK293T cells were transfected with CRISPR-dCas9-VPR plasmids to induce MAPT expression. One day after transfection, luciferase substrates and ITU were added at the indicated concentrations. Two days later, cells were imaged using a bioluminescence live imaging system, or cells were measured longitudinally for 3 d. b, Bioluminescence imaging enabled longitudinal monitoring of 4R-tau in the MAPT-exon-10-tagged HEK293T line at cellular resolution. Respective images were taken 72 h after induction/transfection. Scale bars, 200 μm. c, Histograms of the luminescence signals obtained from three cell dishes without and with MAPT induction. d, RLU of FLuc and NLuc (normalized to DMSO as the vehicle control), and their ratio as a function of increasing the concentrations of the DYRK1A/GSK3A inhibitor ITU in EXSISERSMAPT:10NLuc-11FLuc HEK293T cells. Data are mean ± s.d., n = 3 biological replicates. Selected results of Bonferroni MCT after one-way ANOVA are shown; **P < 0.01, ****P < 0.0001; NS, not significant (full statistical results are provided in Supplementary Table 1). e, Pan-tau and 4R-tau expression was monitored longitudinally through FLuc and NLuc in the double-luciferase EXSISERS HEK293T cell line. Furthermore, 24 h after transfection, ITU at the indicated concentrations, d-luciferin (FLuc substrate) and endurazine (NLuc pro-substrate) were added and measured over a timecourse of 67 h. Data are mean ± s.d. (dotted lines), n = 3 biological replicates. Source data are available online.
