Sigmaplot ver 12

Descriptive statistical measures (mean, median, standard deviation, range, minimum, maximum, and interquartile range) for different soil factors were determined using Sigmaplot ver. 12.5.
Several biodiversity indicators were measured such as Shannon′s and Simpson′s diversity indices, species richness, and evenness were extracted using PC-ORD ver. 5 software [58 ]. Hills numbers were calculated using the R program.
To clarify the relationship between environmental factors, especially soil factors, the study area was divided into three areas: the beginning of the Wadi (stands 1 to 7), the middle (stands 8 to 14), and the end of the Wadi (stands 15 to 20). A detrended canonical correspondence analysis (DCCA) was performed using CANOCO ver. 4.5 and CanoDraw ver. 4.1 [59 (link)].

SigmaPlot Ver.12.5 statistical software was used for all statistical analyses. Larval development assay (LDA; Figures 1, 4E and Supplementary Figure 1) two-way ANOVA (multiple pairwise comparisons, Bonferroni). Total number of pupae (Figure 1B.II) were analyzed with two-way ANOVA (multiple pairwise comparison, Dunn’s). Immunoreactivity (Figure 2C) was analyzed with t-test (Mann–Whitney Rank Sum). Ecdysteroid titers (Supplementary Figures 4A,B) were analyzed with two-way ANOVA (multiple pairwise comparison, Holm-Sidak). Calcium spikes (Figure 4D and Supplementary Figure 4C) were quantified using FIJI¹ and analyzed using two-way ANOVA (all pairwise comparison, Holm-Sidak). Starvation resistance assay (SRA; Figures 5A–C and Supplementary Figures 5A–E) were analyzed with Kaplan–Meyer survival analysis (multiple pairwise comparisons, Holm-Sidak). PER (Figure 5D), lipid content (Figure 5G), protein content (Supplementary Figure 5F), and wing measurements (Figures 5F,H) were analyzed with one-way ANOVA (Holm-Sidak).

Mann–Whitney U rank sum tests followed by the calculation of two-tailed p values were used for determining the significance between groups. SigmaPlot (ver.12.5) was used for graphing and statistical analysis.

Data are presented as mean ± SEM for n arteries obtained from N different animals. Often, only one artery was examined per animal, due to the lengthy duration of the experimental protocol. For each artery, functional responses to pharmacologic agents were first obtained under control conditions and then in the presence of a stated treatment (e.g., 1 μmol/L phenylephrine exposure). Each vessel thus served as its own control for a given manipulation. The difference in the magnitude of response to a given agent under control and treatment conditions (i.e., R2 ‐ R1) was then calculated and presented graphically, as described in Figure 1B. In some cases, a calculated set of differences was analyzed using a one sample t‐test (SigmaPlot ver12) to determine if the mean was statistically different than zero (see Figs. 3, 6B and 8C). For other data, a statistically significant difference for the same stimulus under two experimental conditions was evaluated, using either a Student's unpaired t‐test (see Fig. 8B) or a one‐way ANOVA, followed by a Tukey post hoc test. A calculated P < 0.05 was taken to signify statistical significance.

To perform the statistical analyses on the experimental material, SigmaPlot ver. 12 (SigmaStat, USA) was used. The experimental results were expressed as the mean ± standard deviation (mean ± SD) and statistical significance between groups was determined by using one-way ANOVA followed by Tukey’s post hoc analysis. Values of P < 0.05 were considered to be statistically significant.