Igor pro

Statistical significance was determined by two-tailed unpaired Student’s t test and two-tailed heteroscedastic Student’s t test for Seahorse assay. One-sided t test was used for animal studies, in addition to log-rank test for the Kaplan-Meier plots. GraphPad Prism and Igor Pro were used for calculations. Differences were considered to be statistically significant if P < .05.

Statistics were performed with Cell Quest Pro software (BD Bioscience), HEKA FitMaster v2x53 and IgorPro for patch-clamp transient currents and GraphPad Prism for analysis of dose-response curves and Student’s t-test, with P* < 0.05, P** < 0.01, and P*** < 0.001. The combination index (CI) and dose reduction index (DRI) were determined with CompuSyn1.0 software using the Chou-Talalay method [46 (link)]. For primary AML cells, the coefficient of drug interaction (CDI) was calculated to assess the synergistic inhibitory effect of the drug combination (CDI = AB/(A × B)) [47 (link)].

Data were analyzed using custom software written in Igor Pro (Wavemetrics, Lake Oswego, Oregon) and were plotted with Igor Pro and Prism 5 (Graphpad, La Jolla, California). Data sets were tested for normality using Kolmogorov-Smirnov normality tests. Statistical significance between conditions was determined using paired two-tailed Student's t tests. In order to evaluate the fidelity of phase locking for evoked spiking, we converted individual spike times, with time = 0 coinciding to stimulus onset, to unit vectors with appropriate phase angle for the given stimulus and then calculated vector strength (r) [31] (link).

Electrophysiological recordings were obtained from the C. elegans neuromuscular junctions of immobilized and dissected adult worms as previously described (Richmond, 2009 (link)). Ventral body wall muscle recordings were acquired in whole-cell voltage-clamp mode (holding potential, −60 mV) using an EPC-10 amplifier, digitized at 1 kHz. Evoked responses were obtained using a 2 ms voltage pulse applied to a stimulating electrode positioned on the ventral nerve cord anterior to the recording site. For multiple stimulations, a five pulse train was delivered at 20 Hz. The 5 mM Ca²⁺ extracellular solution consisted of 150 mM NaCl, 5 mM KCl, 5 mM CaCl₂, 4 mM MgCl₂, 10 mM glucose, 5 mM sucrose, and 15 mM HEPES (pH 7.3,~340 mOsm). The patch pipette was filled with 120 mM KCl, 20 mM KOH, 4 mM MgCl₂, 5 mM (N-tris[Hydroxymethyl] methyl-2-aminoethane-sulfonic acid), 0.25 mM CaCl₂, 4 mM Na²ATP, 36 mM sucrose, and 5 mM EGTA (pH 7.2,~315 mOsm). Data were obtained using Pulse software (HEKA. Subsequent analysis and graphing was performed using mini analysis (Synaptosoft), Igor Pro and Prism (GraphPad).

Experimental data were analyzed and presented using Igor Pro and Prism (GraphPad Software, San Diego, CA). For statistical significance, we tested the normality of the data distribution with the Kolmogorov–Smirnov test with the Dallal-Wilkinson-Lillie correction for corrected p values using Prism 5. If a dataset passed the normality test, we used the unpaired Student’s t-test; for all other datasets we used the non-parametric Mann-Whiney U test. For statistical significance of multiple groups, we used a two-way ANOVA. For comparing slopes of lines, we used the linear regression test. Data collected as raw values are shown as mean ± S.E.M. or mean ± S.D. Details of statistical methods are reported in the text. For all analyses, p values < 0.05 were considered significant.

Any statistical packages can be employed for statistical tests. We have utilized commercially available packages including GraphPad Prism (RRID:SCR_002798), Igor Pro and MATLAB. R is widely used for statistical computing software (RRID:SCR_001905) as it is freely available, and it runs on Windows, MacOS and a wide variety of Unix platforms. We have developed a practical R script library for basic biostatistics: RStatisticalTests [16 (link)]. In this library, TwoSampleTest.R is used for parametric and non-parametric comparisons between two independent groups. OneWayANOVA.R is used for parametric and non-parametric comparisons between multiple groups, followed by post-hoc comparisons.

All figures presented were analyzed and graphed in GraphPad Prism 8 or IgorPro software unless stated otherwise. Data shown as mean ± SD. Two group comparisons were carried by Student’s unpaired t test, and percentage comparison was done by χ² test; p < 0.05 was considered significant. N = 3 sheep (two to six weeks post-OD) were used for electrophysiology and Ca²⁺ imaging while another cohort of N = 3 sheep were used for immunohistochemistry.