Each result was repeated at least three times and analyzed by GraphPad prism 6.0 statistical software.
Prism 6.0 statistical software
Manufactured by GraphPad
Sourced in United States
Prism 6.0 is a statistical software package developed by GraphPad. It provides tools for data analysis, curve fitting, and graph creation.
Automatically generated - may contain errors
8 protocols using prism 6.0 statistical software
1
Robust Statistical Analysis Protocol
2
Dexamethasone Inhibits TNFα-Induced CXCL8 Release
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Nasal epithelial cells were treated with dexamethasone for 45 min, followed by TNFα (10 ng/mL) stimulation overnight. The ability of dexamethasone to inhibit TNFα-induced CXCL8 release was determined in cell medium by sandwich ELISA according to the manufacturer’s instructions (R&D Systems, Minneapolis, MN, USA). IC50 of dexamethasone on CXCL8 production (Dex-IC50), calculated using Prism® 6.0 statistical software (GraphPad, San Diego, CA, USA), was used as a marker for corticosteroid sensitivity. In addition, BEAS-2B cells were treated overnight with FITC-conjugated dexamethasone (Molecular Probes, Eugene, OR; 10−6 M). The nuclear fraction was prepared by 10 min incubation with hypotonic buffer (Epigentec, Farmingdale, NY, USA), followed by pulse vortexing. dexamethasone’s ability to translocate into the nucleus was evaluated by FITC-dexamethasone detected in the nuclei using a fluorescent plate reader.
3
Student's t-test for Mutant Evaluation
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All the experimental data were subjected to Student's t-test (P ≤ 0.05) for comparative evaluation of changes in the mutant and the WT both under control and stress conditions using graphpad prism 6.0 statistical software (www.graphpad.com ).
4
Comprehensive Statistical Analysis Protocol
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All independent experimental data were expressed as mean ± standard deviation (SD). Statistical analysis was carried out by using Graphpad prism 6.0 statistical software Inc (Lajolla, USA). P values were calculated by ANOVA and Bonferroni test (more than two groups) or T test (two groups). When P value < 0.05, the results are considered to be statistical.
5
FRAP Analysis of PUFA-Treated PBECs
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PBECs in Ibidi µ-Slide 8-well microscopy chambers were treated with PUFAs, washed three times with warm Hank´s balanced salt solution and incubated for 20 min with Vybrant DiI reagent. After three 5 min washes, 300 µl of warm HBSS were added to each well.
Fluorescence recovery after photobleaching (FRAP) was analysed with a Leica TCS SP5 AOBS inverted confocal microscope (Leica, Germany) with an excitation wavelength of 568 nm. Bleaching was carried out in the region of interest (ROI) at 488 nm for 1 s.
Images were analysed with LAS AF lite software (Leica, Germany) and the extracted data processed using the double normalisation method (Kenworthy, 2007).
The post-bleaching portion of fractional fluorescence recovery curves over time were fitted under an exponential decay function, using the Prism 6.0 statistical software (GraphPad, USA), to obtain the half-time of recovery (t½) parameter. The diffusion coefficient (D) was determined with the following equation (Yamamoto and Ando, 2013) .
Where ω is the radius of the focused laser beam.
Fluorescence recovery after photobleaching (FRAP) was analysed with a Leica TCS SP5 AOBS inverted confocal microscope (Leica, Germany) with an excitation wavelength of 568 nm. Bleaching was carried out in the region of interest (ROI) at 488 nm for 1 s.
Images were analysed with LAS AF lite software (Leica, Germany) and the extracted data processed using the double normalisation method (Kenworthy, 2007).
The post-bleaching portion of fractional fluorescence recovery curves over time were fitted under an exponential decay function, using the Prism 6.0 statistical software (GraphPad, USA), to obtain the half-time of recovery (t½) parameter. The diffusion coefficient (D) was determined with the following equation (Yamamoto and Ando, 2013) .
Where ω is the radius of the focused laser beam.
6
Statistical Analysis of Experimental Data
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Graphpad prism 6.0 statistical software (GraphPad Software, San Diego, California, USA) and SPSS 20.0 (IBM SPSS software, Armonk, New York, USA) were used for statistical analysis and graphing, respectively. All data are expressed as the mean ± SEM. Comparisons between the study groups were done using a one-way analysis of variance test. Post hoc analysis was done using Bonferroni’s method. A probability level of P < 0.05 was considered statistically significant.
7
Binding Activity Preservation Analysis
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Preservation of binding activity upon labelling with IR700 was certified through side-by-side flow-cytometric titration analysis of labeled and unlabeled mAbs. Serially diluted antibody was incubated with 5 × 105 HCT116 colon carcinoma cells (which are AC133 positive) and 5 × 105 p53-deficient HCT116 colon carcinoma cells (which are AC133 negative) suspended in 100 μl FACS buffer (PBS with 0.5% BSA and 2 mM EDTA) for 15 min. After washing, the cells were incubated with 1.5 μg anti-mouse PE-conjugated F(ab')2 fragment (Dianova) in 100 μl FACS buffer for 20 min. Samples were then washed twice and analyzed on a BD FACSVerseTM flow cytometer (BD Bioscience). Statistical analyses were performed using GraphPad Prism 6.0 statistical software (GraphPad Software Inc.).
8
Histological Features Comparison
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Comparisons of histological features among the groups were performed using the Kruskal-Wallis and Dunn's post hoc tests, using Graphpad Prism 6.0 Statistical Software (GraphPad Software Inc., San Diego, USA). Differences were considered significant when p < 0.05.
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