Prism 5 9

We used Imaging J, LabChart 8, FlowJo, Zeiss ZEN pro, Nikon NIS-ELEMENTS-BR for data analysis. We used GraphPad Prism 5/9 for statistical analysis. The results from each study were presented as mean ± SEM. Detailed statistical approaches were described in Figure legends. Differences between means at a level of p ≤ 0.05 were considered to be significant.

For statistical analysis, the Prism 5–9 software (GraphPad https://www.graphpad.com/scientific-software/prism/ (accessed on 10 January 2021)) was used. The data were tested for normal distribution with the Kolmogorov–Smirnov test. For comparison of two groups, an unpaired t-test was performed. For comparison of more than two groups, one-way analysis of variance (ANOVA) was used with Tukey’s or Bonferroni’s multiple comparison test as a post hoc test. Correlation analysis was conducted using Pearson’s correlation test in GraphPad Prism. The data were depicted as mean and standard error of the mean (SEM) or standard deviation (SD) with a 95% confidence interval as indicated in the respective figure legends. P-values of p < 0.0001 and p < 0.001 were considered extremely significant (**** or ***), p < 0.01 very significant (**) and p < 0.05 significant (*).

Purified truncated Mcp was incubated with different concentrations (0, 5, 10, 15, 20 or 25 μM) of As(III), As(V) or Pi at room temperature for 1 h. Tryptophan fluorescence was monitored between 290 and 390 nm with a fluorescence spectrophotometer (PerkinElmer, Massachusetts, USA). The K_D value was calculated by GraphPad Prism 59 (link).