Shortened TDP-43 isoforms suppress endogenous TDP-43 expression. (A) Schematic representation of alternative splicing isoforms of TARDBP. Black or gray boxes indicate coding or untranslated regions, respectively. The pale gray box with a dotted outline represents the region that becomes exon 7 through alternative splicing. Orange and blue lines indicate alternative donor and acceptor sites, respectively. Alternative splicing of TARDBP generates TDP-MPs by excluding part of exon 6 from the TDP-FL, resulting in unique C-terminal sequences (light blue, purple, or green circle with red outlines). The red box marks the siRNA target site specific to TDP-FL. (B) WB analysis of HEK293T cells overexpressing TDP-MPs fused to Venus (MPs-Venus). Quantification of endogenous TDP-FL (FL-endo) levels is normalized to β-actin (n = 3 for each group). Controls include no transfection (NoTF Ctr) and empty vector (Vector Ctr). Data represent the mean ± SEM. Statistical significance was evaluated using one-way ANOVA followed by Dunnett’s test. *P < 0.05, **P < 0.01, and ***P < 0.001 compared with the NoTF Ctr group. (C) Immunocytochemical images of HeLa cells overexpressing FLAG-tagged MP20 (127) or MP18 (127) (FLAG-MPs). Endogenous TDP-43 was detected using an antibody against Gly400 (G400), absent in MPs (green). FLAG (magenta) and Hoechst (blue). Arrowheads indicate FLAG-positive cells. Scale bars: 20 μm. Source data are available for this figure: SourceData F1.
Figure 1.

Shortened TDP-43 isoforms suppress endogenous TDP-43 expression. (A) Schematic representation of alternative splicing isoforms of TARDBP. Black or gray boxes indicate coding or untranslated regions, respectively. The pale gray box with a dotted outline represents the region that becomes exon 7 through alternative splicing. Orange and blue lines indicate alternative donor and acceptor sites, respectively. Alternative splicing of TARDBP generates TDP-MPs by excluding part of exon 6 from the TDP-FL, resulting in unique C-terminal sequences (light blue, purple, or green circle with red outlines). The red box marks the siRNA target site specific to TDP-FL. (B) WB analysis of HEK293T cells overexpressing TDP-MPs fused to Venus (MPs-Venus). Quantification of endogenous TDP-FL (FL-endo) levels is normalized to β-actin (n = 3 for each group). Controls include no transfection (NoTF Ctr) and empty vector (Vector Ctr). Data represent the mean ± SEM. Statistical significance was evaluated using one-way ANOVA followed by Dunnett’s test. *P < 0.05, **P < 0.01, and ***P < 0.001 compared with the NoTF Ctr group. (C) Immunocytochemical images of HeLa cells overexpressing FLAG-tagged MP20 (127) or MP18 (127) (FLAG-MPs). Endogenous TDP-43 was detected using an antibody against Gly400 (G400), absent in MPs (green). FLAG (magenta) and Hoechst (blue). Arrowheads indicate FLAG-positive cells. Scale bars: 20 μm. Source data are available for this figure: SourceData F1.

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