sod1 g93a cat 26401 constructs  (Addgene inc)

Journal: Biomolecules

Article Title: Transcriptomic Analysis Reveals the Beneficial Effects of Spermidine in an ALS Mouse Model

doi: 10.3390/biom16040566

Figure Lengend Snippet: Transcriptomic analysis revealed deregulated genes and pathways in spinal cord and skeletal muscle of SOD1-G93A mice. ( a , b ) Volcano plot of differential gene expression in SOD1-G93A mice versus control mice using the Wald test for spinal cord (SC) and gastrocnemius (GNM) tissues. The statistically significant up-regulated and down-regulated genes are shown in red and blue respectively. ( c , d ) Heatmap of differentially expressed genes in spinal cord and gastrocnemius, respectively, between SOD1-G93A and control mice. Each row of the heatmap represents the z-score-transformed DESeq2 values of one differentially expressed gene across all samples. The row z-score is represented by a color code (blue, down-regulated genes; red, up-regulated genes). ( e , f ) Dotplot of significant pathways derived from gene set enrichment analysis (GSEA) for SC and GNM tissue. The colors represent the p -value adjusted by Benjamini–Hochberg correction. The dot dimension represents the number of DEGs for that pathway. NES, normalized enrichment score. ( g ) Venn diagram showing the number of differentially expressed genes (DEGs) in GNM and SC. Common DEGs are indicated in the intersection. ( h ) Comparative dotplot of common deregulated pathways between tissues.

Article Snippet: SOD1-G93A mice (B6.Cg-Tg(SOD1 G93A)1Gur/J) were obtained from the Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: Gene Expression, Control, Transformation Assay, Derivative Assay

Journal: Biomolecules

Article Title: Transcriptomic Analysis Reveals the Beneficial Effects of Spermidine in an ALS Mouse Model

doi: 10.3390/biom16040566

Figure Lengend Snippet: Validation of RNA-seq analysis in spinal cord and gastrocnemius of SOD1-G93A mice. ( a , b ) RT-qPCR on two deregulated genes from spinal cord (SC) and gastrocnemius (GNM) RNA-seq analysis. Data are represented as mean ± SEM with 2 −ΔΔCt method for n = 3 mice per experimental group. Data were normalized using Gapdh and beta-2-microglobulin (B2m) as housekeeping genes for SC and GNM respectively. Unpaired t -tests were used; * p < 0.05, ** p < 0.01, *** p < 0.001. ( c , d ) Spearman’s correlation test between 2 −ΔCt values and DESeq2 counts from RT-qPCR and RNA-seq analysis, respectively. p -value and Spearman’s r coefficient are displayed in the graphs.

Article Snippet: SOD1-G93A mice (B6.Cg-Tg(SOD1 G93A)1Gur/J) were obtained from the Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: Biomarker Discovery, RNA Sequencing, Quantitative RT-PCR

Journal: Biomolecules

Article Title: Transcriptomic Analysis Reveals the Beneficial Effects of Spermidine in an ALS Mouse Model

doi: 10.3390/biom16040566

Figure Lengend Snippet: Transcriptomic analysis of spinal cord and skeletal muscle of SOD1-G93A and SOD1-G93A treated with SPD. ( a , b ) Volcano plot of differentially expressed genes from spinal cord (SC) and gastrocnemius (GNM) tissues of SOD1-G93A and SOD1-G93A mice treated with SPD (SOD1-G93A SPD). The statistically significant up-regulated and down-regulated genes are shown in red and blue respectively, while grey points represent not significant genes. ( c ) Heatmap of the 10 genes whose expression is reverted in the WT vs. ALS and ALS vs. ALS + SPD comparisons for SC. ( d , e ) Venn diagram and heatmap of 3590 common genes between WT vs. ALS and ALS vs. ALS + SPD for GNM. In Venn diagram are represented DEGs in a red circle for ALS vs. CTR comparison, while in a blue circle for ALS SPD vs. ALS comparison. Each row of the heatmap represents the z-score-transformed DESeq2 values of one differentially expressed gene (DEG) across all samples. The row z-score is represented by a color code (blue, down-regulated genes; red, up-regulated genes). ( f , g ) Dotplot of significant pathways derived from gene set enrichment analysis (GSEA) for SC and GNM tissue respectively. The colors represent the p -value adjusted by Benjamini–Hochberg correction. The dot dimension represents the number of DEGs for that pathway. NES, normalized enrichment score.

Article Snippet: SOD1-G93A mice (B6.Cg-Tg(SOD1 G93A)1Gur/J) were obtained from the Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: Expressing, Comparison, Transformation Assay, Derivative Assay

Journal: Biomolecules

Article Title: Transcriptomic Analysis Reveals the Beneficial Effects of Spermidine in an ALS Mouse Model

doi: 10.3390/biom16040566

Figure Lengend Snippet: Validation of some DEGs in spinal cord tissue of SOD1-G93A mice after SPD treatment. ( a – d ) RT-qPCR of deregulated genes from RNA-seq analysis for spinal cord. Data are represented as mean ± SEM using expression levels measured using 2 −ΔΔCt method for n = 3 mice per experimental group. Data were normalized using Gapdh as housekeeping gene. One-way ANOVA, followed by Tukey’s post hoc test, was used; * p < 0.05, ** p < 0.01, *** p < 0.001. ( e – h ) Spearman’s correlation test was used to assess a correlation between 2 −ΔCt values and DESeq2 counts from RNA-seq analysis. p -value and Spearman’s r coefficient are displayed in the graphs.

Article Snippet: SOD1-G93A mice (B6.Cg-Tg(SOD1 G93A)1Gur/J) were obtained from the Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: Biomarker Discovery, Quantitative RT-PCR, RNA Sequencing, Expressing

Journal: Biomolecules

Article Title: Transcriptomic Analysis Reveals the Beneficial Effects of Spermidine in an ALS Mouse Model

doi: 10.3390/biom16040566

Figure Lengend Snippet: Validation of genes related to mitochondrial and ribosomal protein pathways emerging from GSEA in gastrocnemius tissue of SOD1-G93A mice after SPD treatment. ( a – c , g – i ) RT-qPCR of deregulated genes from RNA-seq analysis of gastrocnemius. Data are mean ± SEM represented using expression levels measured using 2 −ΔΔCt method for n = 3 mice per experimental group. Data were normalized using beta-2-microglobulin (B2m) as housekeeping gene. One-way ANOVA, followed by Tukey’s post hoc test, was used; * p < 0.05, ** p < 0.01. ( d – f , j – l ) Spearman’s correlation test was used to assess a correlation between 2 −ΔCt values and DESeq2 counts from RNA-seq analysis, n = 3 mice per experimental group. p -value and Spearman’s r coefficient are displayed in the graphs.

Article Snippet: SOD1-G93A mice (B6.Cg-Tg(SOD1 G93A)1Gur/J) were obtained from the Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: Biomarker Discovery, Quantitative RT-PCR, RNA Sequencing, Expressing

Journal: Biomolecules

Article Title: Transcriptomic Analysis Reveals the Beneficial Effects of Spermidine in an ALS Mouse Model

doi: 10.3390/biom16040566

Figure Lengend Snippet: Spermidine has a positive effect on mitochondrial bioenergetics in C2C12 cells expressing hSOD1-G93A mutation ( a ) and in NSC34 expressing hTDP43 Q331K mutation ( b ). Individual parameters for basal respiration, maximal respiration, ATP production and spare respiratory capacity were measured. Data are shown as mean ± SEM. One-way ANOVA, with Tukey’s post hoc test, was used. Values significantly different from control (WT) are indicated with # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001. Values significantly different from ALS condition (SOD1 G93A and TDP43 331) are represented with * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: SOD1-G93A mice (B6.Cg-Tg(SOD1 G93A)1Gur/J) were obtained from the Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: Expressing, Mutagenesis, Control

Journal: Biomolecules

Article Title: Transcriptomic Analysis Reveals the Beneficial Effects of Spermidine in an ALS Mouse Model

doi: 10.3390/biom16040566

Figure Lengend Snippet: Body weight and grip strength analyses of SOD1-G93A mice treated with SPD. ( a ) Graph representing the weight of WT, SOD1-G93A and SOD1-G93A + SPD mice at the indicated ages. Data are presented as the means ± SEM; * p < 0.05, ** p < 0.01, *** p < 0.001 for ALS vs. WT comparison. ( b ) Maximal grip strength for WT, SOD1-G93A and SOD1-G93A + SPD mice. Data are presented as the means ± SEM; ** p < 0.01, *** p < 0.001 and **** p < 0.0001 for ALS vs. WT comparison; # p < 0.05 for ALS vs. ALS + SPD comparison; § p < 0.05 and §§ p < 0.01 for ALS SPD vs. WT comparison. Two-way ANOVA Multiple comparisons test was used.

Article Snippet: SOD1-G93A mice (B6.Cg-Tg(SOD1 G93A)1Gur/J) were obtained from the Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: Comparison