<i>Cyp6g1</i> Expression in the Digestive Tissues.

<p><i>Cyp6g1</i> overexpression was achieved in the midgut, Malpighian tubule and the fat body using the GAL4-UAS system. At 0ppm (A, A’), no significant differences were observed between genotypes. At 48ppm imidacloprid the overexpression line responded less as measured by Response Time, End Point RMR values and GLM analysis. All plots display mean RMR or β values with 95% confidence intervals.</p

Imidacloprid Resistance in nAChR Mutants.

<p>Armenia¹⁴, <i>Dα1</i>^<i>M4</i> and <i>Dβ2</i>^<i>L351R</i> were tested using the Wiggle Index. At 0ppm (A, A’), no significant differences were observed between genotypes. As measured by Response Time, End Point RMR values and GLM analysis at 12 and 48ppm imidacloprid (B, B’, C, C’) <i>Dα1</i>^<i>M4</i> responded the least, <i>Dβ2</i>^<i>L351R</i> displayed an intermediate phenotype and Armenia¹⁴ responded the most. All plots display mean RMR or β values with 95% confidence intervals.</p

Spinosad Resistance in nAChR Mutants.

<p>Armenia¹⁴, <i>Dα6</i>^<i>W337*</i> and <i>Dα6</i>^<i>nx</i> were tested using the Wiggle Index at 0ppm (A, A’), no significant differences were observed between genotypes. At 12 ppm (B, B’) both <i>Dα6</i> mutants displayed significantly delayed response times and responded less in the GLM analysis. However, while <i>Dα6</i>^<i>337*</i> differed from Armenia¹⁴ in End Point RMR value, <i>Dα6</i>^<i>nx</i> did not. At 48ppm (C, C’), while Response Times were still delayed in Dα6 mutants, End Point RMR values were not significantly different and GLM analysis indicated that only <i>Dα6</i>^<i>337*</i> responded more than Armenia¹⁴. All plots display mean RMR or β values with 95% confidence intervals.</p

The Wiggle Index.

<p>The Wiggle Index measures total motility in a given well A) Individual wells are cropped out of a sequence of 250.jpeg images B) σ_S values are calculated for each pixel by calculating the standard deviation of all σ_FW values over the entire sequence. C) σ_S values from each pixel are filtered based on a threshold value and then averaged to yield the Wiggle Index (WI) value.</p

Armenia¹⁴ Dose Response Curves.

<p>Armenia¹⁴ was tested using the Wiggle Index at 5 doses of 4 insecticides. Chlorantraniliprole (A) and Imidacloprid (B) each show rapid response times and dose dependent long term RMR values. Ivermectin (C) and Spinosad (D) responded differently, displaying delayed response times and similar long term RMR values. All points correspond to the mean RMR values with 95% confidence intervals. Boxes in each plot indicate values between 0 and 30 minutes magnified in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0145051#pone.0145051.g004" target="_blank">Fig 4</a>.</p

Wiggle Index Output and RMR Calculation.

<p>The Wiggle Index produced heat maps and numeric estimates of total larval motility in each well at a given time. RMR values were calculated by dividing the WI value at time = x from the WI value from the same well at time = 0.</p

Fournier-Leveletal_SupplementaryDataS1

List of accessions and Fitness phenotype in field condition

Fournier-Leveletal_SupplementaryDataS2

SNP markers associated with fitness in multiple field condition

Fournier-Leveletal2016_JEB_DataArchive

Data archive and analysis script in the R programming languag

Fournier-Leveletal2016_Data_Archive

Data and Analysis scripts in the R programming language supporting the article by Fournier-Level et al (2016) PNAS