Journal: Disease Models & Mechanisms

Article Title: Lymphoblastoid cell lines as a model to understand amyotrophic lateral sclerosis disease mechanisms

doi: 10.1242/dmm.031625

Figure Lengend Snippet: SOD1 , TARDBP and FUS mutations lead to changes in soluble protein levels in LCLs . (A-C) Representative immunoblots for total SOD1, TDP-43 and FUS protein expression levels in Ctrl, sALS, SOD1 , TARDBP and FUS LCLs. LDH was used for sample normalization. Densitometric analysis of WB data are performed by ImageJ software. Statistical analysis was carried out on n =4 Ctrl, n =4 sALS, n =3 SOD1 , n =2 TARDBP and n =2 and FUS LCLs. Data are means±s.e.m. and one-way ANOVA followed by Dunnett's multiple comparison test as a post hoc test. * P <0.05 in B, ** P <0.01 in A and *** P <0.001 in C, all relative to Ctrl.

Article Snippet: Statistical analysis was performed by one-way analysis of variance (ANOVA) followed by Dunnett's multiple comparison test as a post hoc test (GraphPad Prism version 5, San Diego, CA, USA).

Techniques: Western Blot, Expressing, Software, Comparison

Journal: Disease Models & Mechanisms

Article Title: Lymphoblastoid cell lines as a model to understand amyotrophic lateral sclerosis disease mechanisms

doi: 10.1242/dmm.031625

Figure Lengend Snippet: SOD1 , TARDBP and FUS mutations showed protein relocalization in LCLs. (A-C) Representative immunoblots for nuclear and cytoplasmic SOD1, TDP-43 and FUS protein expression levels in Ctrl, sALS, SOD1 , TARDBP and FUS LCLs. Histone H1 was used for nuclear sample normalization; LDH was used for cytoplasm sample normalization. Densitometric analysis of WB data are performed by ImageJ software. Statistical analysis was carried out on n =4 Ctrl, n =4 sALS, n =3 SOD1 , n =2 TARDBP and n =2 and FUS LCLs. Data are means±s.e.m. and one-way ANOVA followed by Dunnett's multiple comparison test as a post hoc test. * P <0.05 in C (cytoplasm), ** P <0.01 in C (nucleus) and *** P <0.001 in A and B (cytoplasm), all relative to Ctrl.

Article Snippet: Statistical analysis was performed by one-way analysis of variance (ANOVA) followed by Dunnett's multiple comparison test as a post hoc test (GraphPad Prism version 5, San Diego, CA, USA).

Techniques: Western Blot, Expressing, Software, Comparison

Journal: Disease Models & Mechanisms

Article Title: Lymphoblastoid cell lines as a model to understand amyotrophic lateral sclerosis disease mechanisms

doi: 10.1242/dmm.031625

Figure Lengend Snippet: SOD1 , TARDBP and FUS mutations showed differences in mitochondrial dynamics in LCLs. Representative immunoblots for mitochondrial fusion protein (OPA1 and MFN1), and for mitochondrial and cytoplasm fission protein (Drp1), expression levels in Ctrl, sALS, SOD1 , TARDBP and FUS LCLs (A-C). COX4 was used for mitochondrial sample normalization; GADPH was used for cytoplasm sample normalization. (D) Representative immunoblot for cytochrome c release. The y -axis represents the relative amount of cytochrome c to either Cox4 (white bars) or GAPDH (grey bars). Densitometric analysis of WB data are performed by ImageJ software. Statistical analysis was carried out on n =4 Ctrl, n =4 sALS, n =3 SOD1 , n =2 TARDBP and n =2 and FUS LCLs. Data are means±s.e.m. and one-way ANOVA followed by Dunnett's multiple comparison test as a post hoc test. * P <0.05 in A and C (cytoplasm), all relative to Ctrl. C, cytoplasm; M, mitochondria.

Article Snippet: Statistical analysis was performed by one-way analysis of variance (ANOVA) followed by Dunnett's multiple comparison test as a post hoc test (GraphPad Prism version 5, San Diego, CA, USA).

Techniques: Western Blot, Expressing, Software, Comparison

Journal: Disease Models & Mechanisms

Article Title: Lymphoblastoid cell lines as a model to understand amyotrophic lateral sclerosis disease mechanisms

doi: 10.1242/dmm.031625

Figure Lengend Snippet: SOD1 , TARDBP and FUS mutations showed differences in mitochondrial respiration in LCLs. Normalized OCR measured by Seahorse XF-24 Bioanalyzer in LCLs of healthy controls, sALS and mutated patients (A). Basal cellular oxygen consumption rate (bOCR) increased significantly (** P <0.01; *** P <0.001) in all patients carrying mutations in SOD1 , TARDBP and FUS . sALS patients showed a bOCR like that of Ctrl (B). Mitochondrial oxygen consumption (mOCR) increased significantly in all mutated patients (*** P <0.001) and in sALS (** P <0.01) compared to Ctrl (C). No significant differences were reported in ATP-synthase-coupled mOCR and ATP-synthase-uncoupled mOCR (D,E). The spare respiratory capacity (SRC) was diminished in all patients (mutated and sporadic) compared to healthy controls (*** P <0.001) (F). Data are means±s.e.m. and one-way ANOVA followed by Dunnett's multiple comparison test as a post hoc test.

Article Snippet: Statistical analysis was performed by one-way analysis of variance (ANOVA) followed by Dunnett's multiple comparison test as a post hoc test (GraphPad Prism version 5, San Diego, CA, USA).

Techniques: Comparison

Journal: Disease Models & Mechanisms

Article Title: Lymphoblastoid cell lines as a model to understand amyotrophic lateral sclerosis disease mechanisms

doi: 10.1242/dmm.031625

Figure Lengend Snippet: SOD1 , TARDBP and FUS mutations showed differences in glycolytic flux in LCLs. Injections of glucose, oligomycin and 2-deoxy-glucose (2-DG) were carried out to determine the glycolytic flux ECAR specific, the glycolytic capacity and the glycolytic reserve between ALS patients and control LCLs, respectively (A). The glycolytic flux ECAR specific was downregulated by about 32% in SOD1 mutated patients compared with Ctrl (** P <0.01) (B); no significant differences were reported in the glycolytic capacity and glycolytic reserve (C,D). Data are means±s.e.m. and one-way ANOVA followed by Dunnett's multiple comparison test as a post hoc test.

Article Snippet: Statistical analysis was performed by one-way analysis of variance (ANOVA) followed by Dunnett's multiple comparison test as a post hoc test (GraphPad Prism version 5, San Diego, CA, USA).

Techniques: Control, Comparison