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[Preprint]. 2024 Jun 29:2024.06.26.600814. [Version 1] doi: 10.1101/2024.06.26.600814

Figure 2. Inclusion of cryptic exon precedes caspase 3 activation leading to cleavage of tau in human neurons deficient in TDP-43.

Figure 2.

(A) RT-PCR analysis of TDP-43 cryptic exon targets from day 8, 10, 12, 14 and 16 i3Neurons showing cryptic exon expression begins as early as day 10. (B) Immunoblot showing efficient knockdown of TDP-43 and cleaved caspase 3 expression at day 15 in i3Neurons treated with a scrambled RNA lentivirus or lentivirus targeting TDP-43. (C) Representative immunofluorescence images of 14-day-old i3Neurons using antibodies against MAP2 and TDP-43 (scale bar, 20μm). (D) Representative immunofluorescence images of 14-day-old i3Neurons using antibodies against MAP2 and cleaved caspase 3 to show activated caspase 3 in TDP-43 depleted cells (scale bar, 20μm). (E) Representative immunofluorescence images of 14-day-old i3Neurons using antibodies against Tuj1 and caspase-3-cleaved tau (TauC3) showing increased cleaved tau in TDP-43 depleted cells (scale bar, 20μm). (F) Co-immunofluorescence of cleaved caspase 3 and TauC3 showing colocalization in ~50% neurons (scale bar, 20μm).