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Single-nucleus profiling detects a large transcriptional response to Sox11 overexpression. ( a ) Experimental design in which CST neurons of adult male and female mice were retrogradely labeled by nuclear-localized AAV2-Retro-H2B-mGreenlantern, AAV2-Retro-Sox11, and a unique molecular barcode. Fluorescently labeled tissue from each group was dissected after one, two, four, or six weeks and pooled for single-nucleus profiling. ( b ) UMAP visualization shows separation of Sox11-treated nuclei from control and co-clustering of all Sox11 time points. ( c ) MA plots showing the relationship between transcript abundance and differential gene expression at the four time points, comparing Sox11-treated to control nuclei (non-parametric Wilcoxon rank sum test, p_adj < 0.05), which reveal numerous transcripts are significantly up-regulated (green) and down-regulated (red). ( d ) Gene ontology analysis of transcripts that are significantly upregulated in Sox11-treated nuclei shows enrichment for terms such as neuron development and cytoskeletal organization ( e ) Gene ontology analysis of transcripts that are significantly downregulated in Sox11-treated nuclei shows enrichment for terms including ion transport and synaptic signaling (p-values indicated in plots, Benjamini-Hochberg test).

Journal: Scientific Reports

Article Title: Sox11 overexpression restores embryonic pro-growth transcription in mature corticospinal tract neurons

doi: 10.1038/s41598-025-33479-5

Figure Lengend Snippet: Single-nucleus profiling detects a large transcriptional response to Sox11 overexpression. ( a ) Experimental design in which CST neurons of adult male and female mice were retrogradely labeled by nuclear-localized AAV2-Retro-H2B-mGreenlantern, AAV2-Retro-Sox11, and a unique molecular barcode. Fluorescently labeled tissue from each group was dissected after one, two, four, or six weeks and pooled for single-nucleus profiling. ( b ) UMAP visualization shows separation of Sox11-treated nuclei from control and co-clustering of all Sox11 time points. ( c ) MA plots showing the relationship between transcript abundance and differential gene expression at the four time points, comparing Sox11-treated to control nuclei (non-parametric Wilcoxon rank sum test, p_adj < 0.05), which reveal numerous transcripts are significantly up-regulated (green) and down-regulated (red). ( d ) Gene ontology analysis of transcripts that are significantly upregulated in Sox11-treated nuclei shows enrichment for terms such as neuron development and cytoskeletal organization ( e ) Gene ontology analysis of transcripts that are significantly downregulated in Sox11-treated nuclei shows enrichment for terms including ion transport and synaptic signaling (p-values indicated in plots, Benjamini-Hochberg test).

Article Snippet: Fig. 2 Fluorescence in situ hybridization confirms upregulation of Sox11, Dpysl3, and Dpysl5 in Sox11-treated animals. ( a ) Experimental design involving retrograde transduction of CST neurons with nuclear-localized AAV2-Retro-H2B-mGreenlantern (H2B-mGl) alone or in combination with AAV2-Retro-Sox11, followed 2 weeks later by in situ hybridization to detect candidate genes. ( b – g ) Coronal sections of cortex with CST nuclei labeled by H2B-mGl (green) and in situ hybridization detection of Sox11 ( b , c ), Dpysl3 ( d , e ), and Dpysl5 ( f , g ) in red.

Techniques: Over Expression, Labeling, Control, Gene Expression

Fluorescence in situ hybridization confirms upregulation of Sox11, Dpysl3, and Dpysl5 in Sox11-treated animals. ( a ) Experimental design involving retrograde transduction of CST neurons with nuclear-localized AAV2-Retro-H2B-mGreenlantern (H2B-mGl) alone or in combination with AAV2-Retro-Sox11, followed 2 weeks later by in situ hybridization to detect candidate genes. ( b – g ) Coronal sections of cortex with CST nuclei labeled by H2B-mGl (green) and in situ hybridization detection of Sox11 ( b , c ), Dpysl3 ( d , e ), and Dpysl5 ( f , g ) in red. Panels ( b , d , f ) show control tissue and ( c , e , g ) show Sox11-overexpressing samples with elevated target expression (arrows). ( h – j ) Quantification of average signal intensity in CST nuclei shows significant increases in Sox11 ( h ), Dpysl3 ( i ), and Dpysl5 ( j ). N > 50 neurons from three replicate animals, **** p < 0.0001, unpaired t-test.

Journal: Scientific Reports

Article Title: Sox11 overexpression restores embryonic pro-growth transcription in mature corticospinal tract neurons

doi: 10.1038/s41598-025-33479-5

Figure Lengend Snippet: Fluorescence in situ hybridization confirms upregulation of Sox11, Dpysl3, and Dpysl5 in Sox11-treated animals. ( a ) Experimental design involving retrograde transduction of CST neurons with nuclear-localized AAV2-Retro-H2B-mGreenlantern (H2B-mGl) alone or in combination with AAV2-Retro-Sox11, followed 2 weeks later by in situ hybridization to detect candidate genes. ( b – g ) Coronal sections of cortex with CST nuclei labeled by H2B-mGl (green) and in situ hybridization detection of Sox11 ( b , c ), Dpysl3 ( d , e ), and Dpysl5 ( f , g ) in red. Panels ( b , d , f ) show control tissue and ( c , e , g ) show Sox11-overexpressing samples with elevated target expression (arrows). ( h – j ) Quantification of average signal intensity in CST nuclei shows significant increases in Sox11 ( h ), Dpysl3 ( i ), and Dpysl5 ( j ). N > 50 neurons from three replicate animals, **** p < 0.0001, unpaired t-test.

Article Snippet: Fig. 2 Fluorescence in situ hybridization confirms upregulation of Sox11, Dpysl3, and Dpysl5 in Sox11-treated animals. ( a ) Experimental design involving retrograde transduction of CST neurons with nuclear-localized AAV2-Retro-H2B-mGreenlantern (H2B-mGl) alone or in combination with AAV2-Retro-Sox11, followed 2 weeks later by in situ hybridization to detect candidate genes. ( b – g ) Coronal sections of cortex with CST nuclei labeled by H2B-mGl (green) and in situ hybridization detection of Sox11 ( b , c ), Dpysl3 ( d , e ), and Dpysl5 ( f , g ) in red.

Techniques: Fluorescence, In Situ Hybridization, Transduction, Labeling, Control, Expressing