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

Journal: Scientific Reports

Article Title: Small-molecule flunarizine increases SMN protein in nuclear Cajal bodies and motor function in a mouse model of spinal muscular atrophy

doi: 10.1038/s41598-018-20219-1

Figure Lengend Snippet: Muscle morphology reveals muscle-specific responses to treatment with flunarizine in SMA mice. ( A) Hematoxylin and eosin staining of the soleus from 11-day-old vehice-treated control mouse compared to vehicle- and flunarizine-treated SMA mice. Scale bar, 50 μm. ( B) Analyses of the fiber size in the soleus, plantaris and tibialis of flunarizine- and vehicle-treated control and SMA mice. Data represent fiber area ± SEM (errors bars) of three (5-day-old, in grey) to ≥5 mice (11-day-old, in black) per experimental group. The flunarizine treatment corrects the atrophy in the plantaris and reduces it in the soleus of SMA mice. The statistical analysis was performed using Kruskal-Wallis test followed by Dunn’s multiple comparison rank test. Wiskers are calculated by the Tukey’s method. The * and # between 5- and 11-day-old mice, respectively. One to three symbols represent p < 0.1, 0.01 and 0.001, respectively. ( C) Analyses of the number of fibers for the three muscles for the same number of mice per group. The treatment prevents the loss of fibers in the soleus of SMA mice. ( D) The snRNP-specific reduction of snRNAs is observed in the soleus, plantaris and tibialis of SMA mice. Total RNA was prepared from tissues of flunarizine- and vehicle-treated controls and SMA mice at 11 days of age (3 mice per group). The snRNA levels are determined by RT-qPCR and the relative amount is presented as percent of the vehicle-treated controls. Statistical analyses are performed as in Fig. . The treatment with flunarizine has modest effects on snRNA levels in muscles of SMA mice.

Article Snippet: Statistical analyses were conducted using the non-parametric Kruskal Wallis followed by Dunn’s multiple comparison rank test or two-way ANOVA followed by Tukey’s multiple comparisons test (GraphPad).

Techniques: Staining, Control, Comparison, Muscles, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: Small-molecule flunarizine increases SMN protein in nuclear Cajal bodies and motor function in a mouse model of spinal muscular atrophy

doi: 10.1038/s41598-018-20219-1

Figure Lengend Snippet: Flunarizine mitigates the defects at the neuromuscular junctions of SMA mice. ( A) Immunofluorescent analyses of the neuromuscular junctions (NMJs) in the tibialis using anti-SNAP25 and anti-neurofilament light chain antibodies in red and labeled alpha-bungarotoxin (BGTX) staining for the acetylcholine receptors (AChRs) in green for the pre- and postsynaptic NMJs, respectively. Scale bar, 100 μm. ( B) Frequency histograms depict the proportion of neuromuscular junctions in three stages (plaques, perforated structures and pretzels) in three muscles, namely soleus, plantaris and tibialis. The analysis shows that endplates are more elaborated in the soleus and tibialis of SMA mice upon the flunarizine treatment. Statistical test is two-way ANOVA followed by Tukey’s multiple comparisons test. Three mice per group. ( C) Representative motor endplates from the tibialis of controls and SMA mice. Scale bar, 15 μm. ( D) Quantification of the areas of the AChR clusters reveals a significant increase in endplate size in control and SMA mice upon flunarizine treatment. Kruskal-Wallis followed by Dunn’s multiple comparison rank test. Wiskers are calculated by the Tukey ‘s method. Three mice per group.

Article Snippet: Statistical analyses were conducted using the non-parametric Kruskal Wallis followed by Dunn’s multiple comparison rank test or two-way ANOVA followed by Tukey’s multiple comparisons test (GraphPad).

Techniques: Labeling, Staining, Muscles, Control, Comparison

Journal: Nature Biotechnology

Article Title: A proteome-scale map of the SARS-CoV-2–human contactome

doi: 10.1038/s41587-022-01475-z

Figure Lengend Snippet: a , Tables showing statistical details of NF-κB transcriptional reporter activity in the absence and presence of selected viral proteins under unstimulated (top) and TNFα stimulated (bottom) conditions. One-way ANOVA with Dunnett’s multiple comparisons test, n = 3, adjusted P values are shown. b , Table showing statistical details of NF-κB transcriptional reporter activity at different amounts of transfected viral protein-encoded plasmid under unstimulated (left) and TNFα stimulated conditions (right). One-way ANOVA with Dunnett’s multiple comparisons test, n = 3 and n = 6, respectively, adjusted P values are shown. a and b , Raw data and full analysis is shown in Supplementary Table . c , Table showing statistical details of NF-κB transcriptional reporter activity under unstimulated (left), TNFα-stimulated (middle) and NSP14-induced conditions in WT and IKBKG KO HEK293 cells (two-way ANOVA with Dunnett’s multiple comparisons test, n = 3), adjusted P values are shown. d , Representative anti-IKBKG (top) western blot demonstrating levels of IKBKG in WT and three independent IKBKG knockout clones of HEK293 cells relative to actin beta (ACTB) loading controls (bottom). e , Representative anti-hemagglutinin (HA) western blot demonstrating levels of tagged NSP14 protein in NF-κB induction experiments relative to actin beta (ACTB) loading controls (bottom). f , Table showing statistical details of viral replication in wild-type, mock KO and CRISPR KOs of the indicated HuSCI host proteins. Kruskal-Wallis with Dunn’s multiple comparisons test, n = 9. Adjusted P values are shown. g , Cell viability of mock KO and CRISPR KOs of the indicated HuSCI host proteins relative to WT cells. Kruskal-Wallis with Dunn’s multiple comparisons test, n = 3. Adjusted, Fisher’s exact P values are shown. f and g , Raw data, Fisher’s exact P values, and full analysis is shown in Supplementary Table . h , Cell viability and relative replication of icSARS-CoV-2-nanoluciferase in HEK293 cells (left) and Vero E6 cells (right) at different concentrations of remdesivir. The EC50 values shown for each cell line were calculated with a variable slope model. Error bars: standard deviation of the mean, n = 3 biological repeats, full analysis in Supplementary Table .

Article Snippet: Significance of the KO effect was calculated against the mock KO using an ordinary one-way nonparametric ANOVA Kruskal-Wallis with Dunn’s multiple comparisons test using GraphPad Prism v9.

Techniques: Activity Assay, Transfection, Plasmid Preparation, Western Blot, Knock-Out, Clone Assay, CRISPR, Standard Deviation

Journal: Nature Biotechnology

Article Title: A proteome-scale map of the SARS-CoV-2–human contactome

doi: 10.1038/s41587-022-01475-z

Figure Lengend Snippet: a , Relative NF-κB transcriptional reporter activity in unstimulated (left) and TNF-α-stimulated conditions (one-way analysis of variance (ANOVA) with Dunnett’s multiple comparisons test, P = 0.0395 and P = 0.0047, respectively). Error bars represent standard deviation of the mean, n = 3. b , Relative NF-κB transcriptional reporter activity at different amounts of transfected viral protein-encoding plasmid in unstimulated (top) and TNF-α-stimulated conditions (middle) (one-way ANOVA with Dunnett’s multiple comparisons test: * P = 0.0183, **** P < 0.0001 and ** P = 0.0012, respectively). Error bars represent standard deviation of the mean, n = 3 (top) and n = 6 (middle). Representative anti-hemagglutinin (HA) western blot demonstrating levels of tagged viral protein in titration experiments relative to actin beta (ACTB) loading controls (bottom). a and b , Precise P values, biological repeats and n for each test are shown in Extended Data Fig. and Supplementary Table . c , Relative NF-κB transcriptional reporter activity under unstimulated (left), TNF-α-stimulated (middle) and NSP14-induced conditions in wild-type (WT) and three independent IKBKG KO clones of HEK293 cells (two-way ANOVA with Dunnett’s multiple comparisons test). Error bars represent standard deviation of the mean, n = 3. Precise P values, biological repeats and n for each test are shown in Extended Data Fig. . ctrl., control. d , Schematic of viral replication assay (top) and viral replication in wild-type, mock KO and CRISPR KOs of the indicated HuSCI host targets (bottom) (Kruskal–Wallis with Dunn’s multiple comparisons test, * P = 0.031, ** P = 0.0047, *** P = 0.0003, **** P < 0.0001, respectively). Error bars represent standard deviation of the mean, n = 9. Precise P values, biological repeats and n for each test are shown in Extended Data Fig. and Supplementary Table . e , Fluorescence microscopy images showing replication of icSARS-CoV-2-mNeonGreen in infected Vero E6 cells treated with 10 µM AZ1 or solvent (DMSO, dimethylsulfoxide). f , Cell viability and relative replication of icSARS-CoV-2-nanoluciferase in HEK293 cells (left) and Vero E6 cells (right) at different concentrations of AZ1. The EC 50 values were calculated with a variable slope model. Error bars represent standard deviation of the mean, n = 8 biological repeats and full analysis in Supplementary Table .

Article Snippet: Significance of the KO effect was calculated against the mock KO using an ordinary one-way nonparametric ANOVA Kruskal-Wallis with Dunn’s multiple comparisons test using GraphPad Prism v9.

Techniques: Activity Assay, Standard Deviation, Transfection, Plasmid Preparation, Western Blot, Titration, Clone Assay, Control, Viral Replication Assay, CRISPR, Fluorescence, Microscopy, Infection, Solvent