The activity of the CaM protein was assayed as the ability of the protein to activate phosphodiesterase (PDE) (Sigma, St. Louis, MO, USA), which hydrolyses cAMP (Sigma) to 5′-AMP [30] (link). Then, 5′-AMP was broken down into adenosine and orthophosphate (Pi) by 5′-nucleotidase (Sigma), which was measured using spectrophotometry at 660 nm. An MTT Cell Proliferation and Cytotoxicity Assay Kit (Solarbio, Shanghai, China) were used to determine the cytotoxicity of the compound following the manufacturer’s protocol [31] (link). The median lethal concentration (LC50) for the assay was analysed with IBM SPSS Statistics (v.16.0).
Spss statistics v 16.0
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SPSS Statistics (v.16.0) is a software product for statistical analysis. It provides functionalities for data management, analysis, and visualization.
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4 protocols using spss statistics v 16.0
1
CaM Protein Activation and Cytotoxicity Assay
2
Xenopus Oocyte Expression of TcL-VGCC
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Xenopus oocytes were used to express the TcL-VGCC gene, and the whole-cell currents were recorded using electrode voltage-clamp following the methods described in previous works [33] (link). Data acquisition and analysis were performed using Digidata 1440A and pCLAMP 10.2 software (Axon Instruments, Inc., CA, USA) [34] (link). Dose-response curves for the assay were analysed using IBM SPSS Statistics (v.16.0).
3
Mite Toxicity Detection via Slip-Dip
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The Food and Agriculture Organization (FAO)-recommended slip-dip method was used to detect the toxicity of compounds against mites as described previously [13] (link), and the sublethal concentrations (LC30, LC50 and LC70) against mites were determined through a log-probit analysis of bioassay data using IBM SPSS Statistics (v.16.0, Chicago, IL, USA).
4
Temporal Properties and Clinical Measures in PD-ICD
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Statistical methods for clinical variables are detailed in Supplementary Text. The threshold for statistical significance was p<0.05. We used a Pearson's correlation analysis to explore the associations between the temporal properties and the clinical measures in PD-ICD patients, applying a post-hoc Bonferroni correction for multiple comparisons (IBM SPSS Statistics v16.0).
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