multiple unpaired student’s t-tests graphpad prism version 9.0  (GraphPad Software Inc)

Journal: Cell Death & Disease

Article Title: GOLPH3 promotes endotoxemia-induced liver and kidney injury through Golgi stress-mediated apoptosis and inflammatory response

doi: 10.1038/s41419-023-05975-x

Figure Lengend Snippet: A Hepatocytes (FL83B) and renal proximal tubule cells (HK-2) were treated with LPS (1 µg/ml) for different time points as indicated. The protein expression levels of GOLPH3 were assessed by western blot analysis, and the quantification results are shown ( n = 3). B The cells were treated with LPS (1 µg/ml) for 8 h, representative images of immunofluorescence staining of GOLPH3 (red) were analyzed by confocal microscopy and relative GOLPH3 intensity were quantified ( n = 3). Scale bar, 50 µm. C Immunofluorescence staining of GM130 (green) was performed to examine the Golgi morphology. The representative immunofluorescence staining images of GM130 were examined using confocal microscopy. Relative cells with abnormal and fragmented Golgi were quantified ( n = 3). Scale bar, 20 µm. D , E Relative mRNA levels of pro-inflammatory cytokines ( Tnfα, IL-1β , and IL-6 ) were examined by real-time PCR analysis in FL83B and HK-2 cells, respectively; and relative mRNA expression of each gene was normalized to that of GAPDH ( n = 3). The data are presented as mean ± SEM. One-way ANOVA, followed by Bonferroni’s multiple comparisons was used, * p < 0.05 versus Control.

Article Snippet: Statistical significance was determined using one-way analysis of variance (ANOVA), followed by Bonferroni’s multiple comparisons for multiple groups or unpaired two-tailed Student’s t -test to compare two groups (GraphPad Prism 7 Software, v.7.00, La Jolla, CA, USA).

Techniques: Expressing, Western Blot, Immunofluorescence, Staining, Confocal Microscopy, Real-time Polymerase Chain Reaction, Control

Journal: Cell Death & Disease

Article Title: GOLPH3 promotes endotoxemia-induced liver and kidney injury through Golgi stress-mediated apoptosis and inflammatory response

doi: 10.1038/s41419-023-05975-x

Figure Lengend Snippet: FL83B and HK-2 cells were transfected with Con siRNA or GOLPH3 siRNA for 24 h, and then treated with LPS (1 µg/ml) for 8 h. A , B Western blot analysis was performed to evaluate transfection efficiency of GOLPH3 siRNA after 24 h. C , D After transfection and LPS treatment, the Golgi morphology of cells were examined. Representative images of immunofluorescence staining of GM130 (green) and GOLPH3 (red), and the quantification of cells with fragmented Golgi from each group are shown ( n = 3). Scale bar, 20 µm. The data are presented as mean ± SEM. Two-tailed Student’s t -test (in A , B ) and One-way ANOVA, followed by Bonferroni’s multiple comparisons (in C , D ) were used, * p < 0.05 versus Con siRNA alone, and # p < 0.05 vs LPS with Con siRNA.

Article Snippet: Statistical significance was determined using one-way analysis of variance (ANOVA), followed by Bonferroni’s multiple comparisons for multiple groups or unpaired two-tailed Student’s t -test to compare two groups (GraphPad Prism 7 Software, v.7.00, La Jolla, CA, USA).

Techniques: Transfection, Western Blot, Immunofluorescence, Staining, Two Tailed Test

Journal: Cell Death & Disease

Article Title: GOLPH3 promotes endotoxemia-induced liver and kidney injury through Golgi stress-mediated apoptosis and inflammatory response

doi: 10.1038/s41419-023-05975-x

Figure Lengend Snippet: FL83B cells were transfected with Con siRNA or GOLPH3 siRNA for 24 h, and treated with LPS for 8 h. A , B After transfection, relative mRNA levels of Golph3 and pro-inflammatory mediators ( Tnfα, IL-6, Mcp1 , and Nos2 ) were examined using real-time PCR analysis. Relative mRNA expression was normalized to that of GAPDH ( n = 3). C Representative images of TUNEL staining and the quantification of TUNEL-positive cells/HPF using Image J software (NIH) are shown. Recombinant DNase I (5 U/ml) treated cells were used as experimental positive control ( n = 3). Scale bar, 50 µm. D After transfection, the cells were treated with LPS for 1 h, and the protein expression levels of GOLPH3, p-NF-κB p65, p-IκBα, p-AKT, and β-actin (as a loading control) were analyzed by western blot analysis ( n = 3). The data are presented as mean ± SEM. One-way ANOVA, followed by Bonferroni’s multiple comparisons was used, * p < 0.05 versus Con siRNA alone, and # p < 0.05 vs LPS with Con siRNA.

Article Snippet: Statistical significance was determined using one-way analysis of variance (ANOVA), followed by Bonferroni’s multiple comparisons for multiple groups or unpaired two-tailed Student’s t -test to compare two groups (GraphPad Prism 7 Software, v.7.00, La Jolla, CA, USA).

Techniques: Transfection, Real-time Polymerase Chain Reaction, Expressing, TUNEL Assay, Staining, Software, Recombinant, Positive Control, Control, Western Blot

Journal: Cell Death & Disease

Article Title: GOLPH3 promotes endotoxemia-induced liver and kidney injury through Golgi stress-mediated apoptosis and inflammatory response

doi: 10.1038/s41419-023-05975-x

Figure Lengend Snippet: A , B After transfection of GOLPH3 siRNA for 24 h, the HK-2 cells were treated with LPS for 8 h, and examined relative mRNA levels of Golph3 and pro-inflammatory mediators ( Tnfα, IL-6, Mcp1 , and Nos2 ) using real-time PCR analysis. Relative mRNA expression was normalized to that of GAPDH ( n = 3). C Representative images of TUNEL staining and quantification of TUNEL-positive cells/HPF using Image J software (NIH) are shown. Recombinant DNase I (5 U/ml) treated cells were used as experimental positive control ( n = 3). Scale bar, 50 µm. D After transfection, the cells were also treated with LPS for 1 h, and the protein expression levels of GOLPH3, p-NF-κB p65, p-IκBα, p-AKT, and β-actin (as a loading control) were analyzed by western blot analysis ( n = 3). The data are presented as mean ± SEM. One-way ANOVA, followed by Bonferroni’s multiple comparisons was used, * p < 0.05 versus Con siRNA alone, and # p < 0.05 vs LPS with Con siRNA.

Article Snippet: Statistical significance was determined using one-way analysis of variance (ANOVA), followed by Bonferroni’s multiple comparisons for multiple groups or unpaired two-tailed Student’s t -test to compare two groups (GraphPad Prism 7 Software, v.7.00, La Jolla, CA, USA).

Techniques: Transfection, Real-time Polymerase Chain Reaction, Expressing, TUNEL Assay, Staining, Software, Recombinant, Positive Control, Control, Western Blot

Journal: Cell Death & Disease

Article Title: GOLPH3 promotes endotoxemia-induced liver and kidney injury through Golgi stress-mediated apoptosis and inflammatory response

doi: 10.1038/s41419-023-05975-x

Figure Lengend Snippet: RAW 264.7 cells were treated with LPS (1 µg/ml) for different time points as indicated. A After LPS treatment, GOLPH3 expression was examined by western blot analysis ( n = 3). B The cells were transfected with Con siRNA or GOLPH3 siRNA for 24 h and the knockdown efficiency was determined by western blot analysis. C , D The cells were transfected with Con siRNA or GOLPH3 siRNA for 24 h, and subsequently treated with LPS for 8 h. The relative mRNA levels of Golph3 and pro-inflammatory mediators ( Tnfα, IL-6, Mcp1 , and Nos2 ) were determined using real-time PCR analysis. Relative mRNA expression was normalized to that of GAPDH ( n = 3). E After transfection, the cells were treated with LPS for 1 h, and determined the protein expression levels of GOLPH3, p-NF-κB p65, p-IκBα, p-AKT, and β-actin (as a loading control) using western blot analysis ( n = 3). F Cells were transfected with Con siRNA or GOLPH3 siRNA for 24 h and treated with LPS for 8 h, and cell lysates were analyzed by western blotting to measure GOLPH3, iNOS, and COX2 expression. The data are presented as mean ± SEM. One-way ANOVA, followed by Bonferroni’s multiple comparisons (in A , C , D , E , F ) and two-tailed Student’s t -test (in B ) were used, * p < 0.05 versus Con siRNA alone, and # p < 0.05 vs LPS with Con siRNA.

Article Snippet: Statistical significance was determined using one-way analysis of variance (ANOVA), followed by Bonferroni’s multiple comparisons for multiple groups or unpaired two-tailed Student’s t -test to compare two groups (GraphPad Prism 7 Software, v.7.00, La Jolla, CA, USA).

Techniques: Expressing, Western Blot, Transfection, Knockdown, Real-time Polymerase Chain Reaction, Control, Two Tailed Test