Journal: bioRxiv

Article Title: Corticospinal propagation of full-length TDP-43 toxicity drives brain-to-muscle pathology

doi: 10.64898/2026.03.27.714692

Figure Lengend Snippet: A) Schematic representation of the intracortical infusion side of FL TDP-43 (IL, ipsilateral, dashed red line) and vehicle (CL, contralateral, blue line). B) Total volume occupied by pTDP-43 colocalized with NeuN+ cells in the motor cortex four-months post-infusion. Values represent the mean ± SEM (One-way ANOVA followed by Tukey’s post hoc test.) ***p<0.001 vs TDP-43 IL; **p<0.01 vs TDP-43 IL. C) Confocal images showing pTDP-43 (yellow) in NeuN+ cells (red). Scale bar: 30 μm. Dashed red square in the picture correspond to the affected motor cortex site (ipsilateral to the injection side). D) Number of NeuN+ neurons measured within the motor cortex. Values represent the mean ± SEM (One-way ANOVA followed by Tukey’s post hoc test). E-I ) Representative western blot analysis ( E ) and relative quantifications of the full-length TDP-43 ( F ) and pTDP-43 ( G ). The graphs represent the densitometric analyses of TDP-43 and phospho-TDP-43 levels normalized using GAPDH as loading control. Data are expressed as arbitrary densitometric units (ADU). Each bar represents the mean ± SEM of five independent replicates (ANOVA with Tukey’s post hoc test among groups). H-I ) Representative filter retardation assay ( H ) and relative densitometric analysis ( I ) of C-terminal insoluble TDP-43 species. Value represents the mean ± SEM of five independent replicates (ANOVA with Tukey’s post hoc test among groups). Please note that dotted red columns in the graphs always refer to the TDP-43-affected side (i.e., ipsilateral to the injection side for the motor cortex and contralateral to the injection side for the spinal cord and muscle). VEH = vehicle.

Article Snippet: Membranes were blocked for 1 hour in 5% non-fat dry milk (PanReac AppliChem ITW Reagents, Darmstadt, Germany, A0830,0500) diluted in TBS-T buffer [20 mM Tris base, 140 mM NaCl, pH 7.6, and 0.1% Tween 20 (Merck, Darmstadt, Germany, P1379)] and incubated overnight at 4 °C with primary antibodies: anti-GAPDH (Immunological Science, Roma, Italy, MAB-10578, 1:5,000), anti-TDP-43 C-term (Proteintech, Sankt Leon-Rot, Germany, 12892-1-AP, 1:1,000), anti Phospho-TDP43 (Ser409/410) (pTDP-43) (Proteintech, 66318-1-IG, 1:1,000), anti-HSP70/HSC70 (Santa Cruz Biotrechnology, Santa Cruz, CA, USA, sc-24, 1:1,000), anti-SQSTM1/p62 (Merck, P0067, 1:3,000), anti-HSPB8 (Thermo Fisher Scientific, Waltham, MA, USA, PA5-76780, 1:1,000), and anti-BAG3 (Abcam, Cambridge, United Kingdom, ab47124, 1:3,000), anti-LC3 (Merck, L8918, 1:3,000).

Techniques: Injection, Western Blot, Control

Journal: bioRxiv

Article Title: Corticospinal propagation of full-length TDP-43 toxicity drives brain-to-muscle pathology

doi: 10.64898/2026.03.27.714692

Figure Lengend Snippet: A) Schematic representation of the intracortical infusion side of FL TDP-43 (IL, ipsilateral, dashed red line) and vehicle (CL, contralateral, blue line). B) Number of NeuN+ neurons measured within the cervical tract of the spinal cord. Values represent the mean ± SEM (One-way ANOVA followed by Tukey’s test) ^p<0.05 vs TDP-43 CL. C) Total volume occupied by pTDP-43 colocalized with NeuN+ cells in the motor cortex four-months post-infusion. Values represent the mean ± SEM (Kruskall-Wallis followed by Dunn’s post hoc test.) *p<0.05 vs TDP-43 CL and VEH CL. D) Confocal images showing pTDP-43 (yellow) in NeuN+ cells (red). Scale bar: 30 μm. Dashed red square in the picture correspond to the affected spinal cord site (contralateral to the injection side). E-G ) Representative western blot analysis ( E ) and relative quantifications of the C-terminal TDP-43 ( F ) and pTDP-43 ( G ) species. The graphs represent the densitometric analyses of C-terminal and phospho-TDP-43 signals normalized using GAPDH as loading control. Data are expressed as arbitrary densitometric units (ADU). Each bar represents the mean ± SEM of five independent replicates (ANOVA with Tukey’s post hoc test among groups). H-I ) Representative filter retardation assay ( H ) and relative densitometric analysis ( I ) of C-terminal insoluble TDP-43 species. Value represents the mean ± SEM of five independent replicates (ANOVA with Tukey’s post hoc test among groups). §p<0.05 vs TDP-43 IL. Please note that dotted red columns in the graphs always refer to the TDP-43-affected side (i.e., ipsilateral to the injection side for the motor cortex and contralateral to the injection side for the spinal cord and muscle). CL = contralateral to the injection site; IL = ipsilateral to the injection site; VEH = vehicle.

Article Snippet: Membranes were blocked for 1 hour in 5% non-fat dry milk (PanReac AppliChem ITW Reagents, Darmstadt, Germany, A0830,0500) diluted in TBS-T buffer [20 mM Tris base, 140 mM NaCl, pH 7.6, and 0.1% Tween 20 (Merck, Darmstadt, Germany, P1379)] and incubated overnight at 4 °C with primary antibodies: anti-GAPDH (Immunological Science, Roma, Italy, MAB-10578, 1:5,000), anti-TDP-43 C-term (Proteintech, Sankt Leon-Rot, Germany, 12892-1-AP, 1:1,000), anti Phospho-TDP43 (Ser409/410) (pTDP-43) (Proteintech, 66318-1-IG, 1:1,000), anti-HSP70/HSC70 (Santa Cruz Biotrechnology, Santa Cruz, CA, USA, sc-24, 1:1,000), anti-SQSTM1/p62 (Merck, P0067, 1:3,000), anti-HSPB8 (Thermo Fisher Scientific, Waltham, MA, USA, PA5-76780, 1:1,000), and anti-BAG3 (Abcam, Cambridge, United Kingdom, ab47124, 1:3,000), anti-LC3 (Merck, L8918, 1:3,000).

Techniques: Injection, Western Blot, Control