Fig. 7: ASO-GGC (200 nM) rescues pathological polyG-induced genomic damage and cellular senescence. | Nature Communications

Fig. 7: ASO-GGC (200 nM) rescues pathological polyG-induced genomic damage and cellular senescence.

From: ASO therapy rescues NOTCH2NLC GGC repeat expansion-induced genomic damage, 3D chromatin structural abnormalities, and senescence

Fig. 7: ASO-GGC (200 nM) rescues pathological polyG-induced genomic damage and cellular senescence.The alternative text for this image may have been generated using AI.

Immunoblotting analysis (a) and quantitative analysis of NPM1 (b) and p-γH2AX (c) protein levels demonstrating that ASO-GGC treatment significantly reduces their levels in patient-derived GGCexp neurons compared to ASO-ctr, normalized to GAPDH. Neurons were treated at 30 DIV for 2 weeks prior to analysis. Statistical analysis was performed using a two-tailed paired Student’s t test (n = 3 biological replicates). Immunoblotting analysis (d) and quantitative analysis of NPM1 (e) and p-γH2AX (f) protein levels demonstrating that ASO-GGC treatment significantly reduces their levels in patient-derived GGCexp-3DCOs compared to ASO-ctr, normalized to GAPDH. Organoids were treated at 76 DIV for 2 weeks prior to analysis; three organoids were selected per sample for analysis. Statistical analysis was performed using a two-tailed paired Student’s t test (n = 3 biological replicates). Immunofluorescence images (g) and quantitative analysis (h) showing that ASO-GGC treatment reduces the percentage of p-γH2AX-positive cells in patient-derived GGCexp neurons compared to ASO-ctr. Statistical analysis was performed using a two-tailed paired Student’s t test (n = 3 biological replicates). Arrows indicate cells positive for both ubiquitin inclusions and γH2AX foci. Scale bar: 50 μm. Immunofluorescence images (i) and quantitative analysis (j) showing that ASO-GGC treatment reduces the percentage of p-γH2AX-positive cells in patient-derived GGCexp-3DCOs compared to ASO-ctr. Statistical analysis was performed using a two-tailed paired Student’s t test (n = 3 biological replicates). Scale bar: 20 μm. Images of β-galactosidase staining (k) and quantitative analysis (l) showing a significant decrease in the area of β-galactosidase-positive blue precipitates in ASO-GGC-treated patient-derived GGCexp neurons compared to ASO-ctr. Statistical analysis was performed using a two-tailed paired Student’s t test (n = 3 biological replicates). Scare bar: 20 μm. Images of β-galactosidase staining (m) and quantitative analysis (n) showing a significant decrease in the area of β-galactosidase-positive blue precipitates in ASO-GGC-treated patient-derived GGCexp-3DCOs compared to ASO-ctr. Statistical analysis was performed using a two-tailed paired Student’s t test (n = 3 biological replicates). Scare bar: 50 μm. Data are presented as mean ± SD.

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