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

Journal: Oncotarget

Article Title: Losartan improves the therapeutic effect of metronomic cyclophosphamide in triple negative mammary cancer models

doi: 10.18632/oncotarget.27694

Figure Lengend Snippet: Tumor growth assessment: data for each time-point in mm 3 are mean ± SEM. ( A ) M-234p, N = 6/group; Day 21: Control (600.96 ± 93) vs Cy+Los (20.96 ± 9.01) ( P < 0.05); ( B ) M-406, N = 6–7/group; Day 17: Control (1397.00 ± 328.32) vs Cy (372.00 ± 55.25) ( P < 0.01), vs Los (451.07 ± 143.94) ( P < 0.05), vs Cy+Los (123.43 ± 45.71) ( P < 0.001). Kruskal-Wallis multiple comparison test and Dunn’s post-test. Overall survival (Kaplan-Meier), Median Survival (MS): ( C ) M-234p, N = 5–6/group; Control (MS: 34 days); Cy (MS: 47 days); Los (MS: 32 days); Cy+Los (MS: undefined, Day 32: 60% [3/5] complete tumor regressions). Cy+Los vs Control, vs Los, vs Cy ( P < 0.01); ( D ) M-406, N = 6–7/group); Control (MS: 24 days); Cy (MS: 36.5 days); Los (MS: 33 days); Cy+Los (MS: 47 days). Control vs Cy ( P < 0.01), vs Los ( P < 0.01), vs Cy+Los ( P < 0.001); Cy+Los vs Cy ( P < 0.05), vs Los ( P < 0,001). Log-rank Test.

Article Snippet: Data obtained was analyzed using ANOVA and Tukey-Kramer Multiple Comparison tests, Kruskal-Wallis and Dunn’s post-test, and Log-rank tests were used to examine the differences between groups with GraphPad Prism version 3.0 (GraphPad Software, San Diego, CA).

Techniques: Control, Comparison

Journal: Oncotarget

Article Title: Losartan improves the therapeutic effect of metronomic cyclophosphamide in triple negative mammary cancer models

doi: 10.18632/oncotarget.27694

Figure Lengend Snippet: Proliferation: Ki67 + cells/field (median, range). ( A ) M-234p Control vs Cy+Los ( P < 0.01); ( B ) M-406 Control vs Cy+Los ( P < 0.05; ( C ) M-234p and ( D ) M-406, representative images of Control and Cy+Los treated tumors, 1000× magnification. Apoptosis: TUNEL + cells/field (median, range). ( E ) M-234p Control vs Cy+Los ( P < 0.05): ( F ) M-406 N. S; Kruskal-Wallis multiple comparison test and Dunn’s post-test; ( G ) M-234p and ( H ) M-406 representative images of Control and Cy+Los treated tumors, 1000× magnification.

Article Snippet: Data obtained was analyzed using ANOVA and Tukey-Kramer Multiple Comparison tests, Kruskal-Wallis and Dunn’s post-test, and Log-rank tests were used to examine the differences between groups with GraphPad Prism version 3.0 (GraphPad Software, San Diego, CA).

Techniques: Control, TUNEL Assay, Comparison

Journal: Oncotarget

Article Title: Losartan improves the therapeutic effect of metronomic cyclophosphamide in triple negative mammary cancer models

doi: 10.18632/oncotarget.27694

Figure Lengend Snippet: Hematoxylin and eosin (H&E) representative tumor sections from M-234p and M-406, 400×. In both models the behavior was similar. Control group: ( A ) M-234p and ( C ) M-406: capillaries with small endothelial cells with barely stained nuclei and intercellular gaps (yellow arrow), lack of pericytes or cells with structure and staining compatible with pericytes. Cy+Los group: ( B ) M-234p and ( D ) M-406: intra- and peritumoral capillaries with structure and morphology similar to normal tissues. Endothelial cells with defined nuclei provide a continuous uninterrupted lining (yellow arrow), and well defined basal membrane covered with pericytes (red arrow). M-406 magnified section (1000×): vessel with normal vascular morphology. HIF1α expression: HIF1α + cells/field (median, range). ( E ) Control vs Cy ( P < 0.05), ( F ) Control vs Cy ( P < 0.05), vs Cy+Los ( P < 0.05), ( G ) and H ), representative images of Control and Cy+Los treated tumors, 100× magnification. Kruskal-Wallis multiple comparison test and Dunn’s post-test.

Article Snippet: Data obtained was analyzed using ANOVA and Tukey-Kramer Multiple Comparison tests, Kruskal-Wallis and Dunn’s post-test, and Log-rank tests were used to examine the differences between groups with GraphPad Prism version 3.0 (GraphPad Software, San Diego, CA).

Techniques: Control, Staining, Membrane, Expressing, Comparison

Journal: Oncotarget

Article Title: Losartan improves the therapeutic effect of metronomic cyclophosphamide in triple negative mammary cancer models

doi: 10.18632/oncotarget.27694

Figure Lengend Snippet: M-234p: ( A ) CD4 cells, N. S. ( B ) CD8 cells, N. S. ( C ) Treg cells, N. S. ( D ) Th17 cells, N. S. M-406: ( E ) CD4 cells, N. S. ( F ) CD8 cells, N. S. ( G ) Treg cells, ( P = 0.064). ( H ) Th17 cells: Control vs Cy+Los, ( P = 0.0580). Kruskal-Wallis multiple comparison test and Dunn’s post-test.

Article Snippet: Data obtained was analyzed using ANOVA and Tukey-Kramer Multiple Comparison tests, Kruskal-Wallis and Dunn’s post-test, and Log-rank tests were used to examine the differences between groups with GraphPad Prism version 3.0 (GraphPad Software, San Diego, CA).

Techniques: Control, Comparison

Journal: Oncotarget

Article Title: Losartan improves the therapeutic effect of metronomic cyclophosphamide in triple negative mammary cancer models

doi: 10.18632/oncotarget.27694

Figure Lengend Snippet: Lymphocytes/field (median, range). M-234p: ( A ) CD4 + cells, N. S. ( B ) CD8 + cells, N. S. ( C ) Foxp3 + cells: Control vs Los, P < 0.05, vs Cy+Los, ( P < 0.05); ( D – F ) representative images of Control and Cy+Los treated tumors, 100× magnification; M-406: ( G ) CD4 + cells, N. S. ( H ) CD8 + cells, N. S. ( I ) Foxp3 + cells: Control vs Cy+Los, ( P < 0.05); ( J – L ) representative images of Control and Cy+Los treated tumors, 100× magnification. Kruskal-Wallis multiple comparison test and Dunn’s post-test.

Article Snippet: Data obtained was analyzed using ANOVA and Tukey-Kramer Multiple Comparison tests, Kruskal-Wallis and Dunn’s post-test, and Log-rank tests were used to examine the differences between groups with GraphPad Prism version 3.0 (GraphPad Software, San Diego, CA).

Techniques: Control, Comparison

Journal: Oncotarget

Article Title: Losartan improves the therapeutic effect of metronomic cyclophosphamide in triple negative mammary cancer models

doi: 10.18632/oncotarget.27694

Figure Lengend Snippet: Quantification of tumor infiltrating lymphocytes by flow cytometry: M-234p, day 42, Cy vs Cy+Los: ( A ) CD4 cells, N. S. ( B ) CD8 cells, N. S. ( C ) Treg cells, ( P < 0.001). ( D ) Th17 cells, ( P < 0.05). Kruskal-Wallis multiple comparison test and Dunn’s post -test.

Article Snippet: Data obtained was analyzed using ANOVA and Tukey-Kramer Multiple Comparison tests, Kruskal-Wallis and Dunn’s post-test, and Log-rank tests were used to examine the differences between groups with GraphPad Prism version 3.0 (GraphPad Software, San Diego, CA).

Techniques: Flow Cytometry, Comparison

Journal: Oncotarget

Article Title: Losartan improves the therapeutic effect of metronomic cyclophosphamide in triple negative mammary cancer models

doi: 10.18632/oncotarget.27694

Figure Lengend Snippet: αSMA : % of αSMA + area/field (median, range). ( A ) M-234p Control vs Los, ( P < 0.05), vs Cy+Los, ( P < 0.01). ( B ) M-406 Control vs Cy, ( P < 0.01), vs Los ( P < 0.01), vs Cy+Los ( P < 0.001). ( C ) M-234p and ( D ) M-406 representative images of Control and Cy+Los treated tumors, 100× magnification. Kruskal-Wallis multiple comparison test and Dunn’s post-test. Collagen : % of collagen area/field (median, range). ( E ) M-234p Control vs Cy+Los ( P < 0.01). ( F ) M-406 N. S. ( G ) M-234p and ( H ) M-406 representative images of Control and Cy+Los treated tumors, 100× magnification.

Article Snippet: Data obtained was analyzed using ANOVA and Tukey-Kramer Multiple Comparison tests, Kruskal-Wallis and Dunn’s post-test, and Log-rank tests were used to examine the differences between groups with GraphPad Prism version 3.0 (GraphPad Software, San Diego, CA).

Techniques: Control, Comparison