Additional file 8 of DVT: a high-throughput analysis pipeline for locomotion and social behavior in adult Drosophila melanogaster
Creators: name: Mi, Kai
name: Li, Yiqing
name: Yang, Yuhang
name: Secombe, Julie
name: Liu, Xingyin
Publisher: name: figshare
Year: 2024
Subjects: subject: Genetics
subject: FOS: Biological sciences

description: Additional file 8: Supplemental Figures Figure S1 DVT analysis pipeline. A, Traditional analysis pipeline with reID error corrected. B, DVT analysis pipeline by average-by-video value of behavior metrics. C, Average-by-video fly behavior metrics has a strong correlation before and after reID error correction. D, Analysis pipeline for heterogeneous Dro. chamber. Statistical analysis: Pearson correlation were calculated between the average-by-video fly behavior metrics before and after reID error correction of n = 8 videos (4 videos from w1118 flies, and 4 videos from OR flies). Figure S2 Behavior deviation for flies in 2 mm-height chamber. A, Effect size for locomotion behavior deviation of drosophila in 2 mm-height chamber from that in 3 mm-height chamber. A.1 ~ A.6, Scatter plots for track straightness at arena edge of male or female flies in 2 mm-height or 3 mm-height chamber from different experimental replicates. B, Effect size for social behavior deviation of drosophila in 2 mm-height chamber from that in 3 mm-height chamber. Interpretation of the radar plot: The effect size was calculated to illustrate the behavior feature deviation of flies in treatment group chambers from that in control chambers. In the calculation, the mean difference was divided by maximum value of standard deviations of the control group and treatment group to denote the effect size. In this case, the control group was fly behavior features in 3 mm-height chamber and the treatment group was that in 2 mm-height profile. In general, both the mean difference between two groups and the intragroup variance affected the magnitude of effect size. if the magnitude of effect size is smaller than 0.2, it is considered to be a small deviation, an effect size of around 0.5 is considered to be a medium deviation, and an effect size of 0.8 or larger is considered to be a large deviation. In the visualization, fly behavior in 3 mm-height chamber was set as the bench and represented by the bold black circle in the radar plot. Each dot in the plot represented the deviation of features from one experiment replicate. For example, we got six dots at the Track straightness at arena edge, from 3 male-fly experiment replicates and 3 female-fly experiment replicates (A.1 ~ A.6). Dots inner the bench circle indicated a decrease of features from control group. While dots outside the bench circle indicated an increase of features from control group. We also color-annotated the label of statistically significant changed features, red for increasing and green for decreasing with q-value asterisks. Statistical analysis: A modified Cohen’s d was used to calculate the effect size of behavioral deviation for flies in 2 mm-height chamber to that in 3 mm-height chamber from 3 experimental replicates. ANOVA were used to identify statistical significantly changed behavior metrics given experimental replicate and fly gender. False discovery rate (Benjamini/Hochberg FDR) was performed for p-value adjustment. Welch Two Sample t-test was used for comparisons between two groups in A.1 ~ A.6. *P &lt; 0.05, **P &lt; 0.01, ***P &lt; 0.001. Figure S3 Behavior deviation for flies with different density profiles. A, Effect size for locomotion behavior deviation of drosophila in 4-flies chamber from that in 6-flies chamber. B, Effect size for social behavior deviation of drosophila in 4-flies chamber from that in 6-flies chamber. C, Effect size for locomotion behavior deviation of drosophila in 8-flies chamber from that in 6-flies chamber. D, Effect size for social behavior deviation of drosophila in 8-flies chamber from that in 6-flies chamber. Statistical analysis: A modified Cohen’s d was used to calculate the effect size of behavioral deviation for flies with different density profiles from 3 experimental replicates. ANOVA were used to identify statistical significantly changed behavior metrics given experimental replicate and fly gender. False discovery rate (Benjamini/Hochberg FDR) was performed for p-value adjustment. Labels of statistically significant changed features were color-annotated, red for increasing and green for decreasing with q-value asterisks. *P &lt; 0.05, **P &lt; 0.01, ***P &lt; 0.001. Figure S4 Behavior deviation for flies in red dim darkness. A, Effect size for locomotion behavior deviation of drosophila in red dim darkness from that under normal Illumination. B, Effect size for social behavior deviation of drosophila in red dim darkness from that under normal Illumination. Statistical analysis: A modified Cohen’s d was used to calculate the effect size of behavioral deviation for flies in red dim darkness with that in normal Illumination from 3 experimental replicates. ANOVA were used to identify statistical significantly changed behavior metrics given experimental replicate and fly gender. False discovery rate (Benjamini/Hochberg FDR) was performed for p-value adjustment. Labels of statistically significant changed features were color-annotated, red for increasing and green for decreasing with q-value asterisks. *P &lt; 0.05, **P &lt; 0.01, ***P &lt; 0.001. Figure S5 Behavior deviation for flies with different experimental times. A, Effect size for locomotion behavior deviation of flies recorded at ZT06 from that at ZT01. B, Effect size for social behavior deviation of flies recorded at ZT06 from that at ZT01. C, Effect size for locomotion behavior deviation of flies recorded at ZT11 from that at ZT06. D, Effect size for social behavior deviation of flies recorded at ZT11 from that at ZT06. Statistical analysis: A modified Cohen’s d was used to calculate the effect size of behavioral deviation for flies with different experimental times from 3 experimental replicates. ANOVA were used to identify statistical significantly changed behavior metrics given experimental replicate and fly gender. False discovery rate (Benjamini/Hochberg FDR) was performed for p-value adjustment. Labels of statistically significant changed features were color-annotated, red for increasing and green for decreasing with q-value asterisks. *P &lt; 0.05, **P &lt; 0.01, ***P &lt; 0.001. Figure S6 Drosophila behavior feature correlation diagram. Statistical analysis: Multi-level correlation of paired behavior metrics was calculated to build the network. Red eclipses indicate positive correlation and green ones are for negative correlations. Only correlation with FDR q-value &lt; 0.05 was colored. Figure S7 Behavior deviation for flies at the last 5 min in the recorded video from that at the first 5 min. A, Effect size for locomotion behavior deviation for w1118 flies at the last 5 min in the video from that at the first 5 min. B, Effect size for social behavior deviation for w1118 flies at the last 5 min in the video from that at the first 5 min. C, Effect size for locomotion behavior deviation for OR flies at the last 5 min in the video from that at the first 5 min. D, Effect size for social behavior deviation for OR flies at the last 5 min in the video from that at the first 5 min. E, Effect size for locomotion behavior deviation for CS flies at the last 5 min in the video from that at the first 5 min. F, Effect size for social behavior deviation for CS flies at the last 5 min in the video from that at the first 5 min. Statistical analysis: A modified Cohen’s d was used to calculate the effect size of behavioral deviation for flies at the last 5 min in the recorded video compared with that at the first 5 min from 3 experimental replicates. Mixed ANOVA were used to identify statistical significantly changed behavior metrics given experimental replicate, fly gender, and video identity. False discovery rate (Benjamini/Hochberg FDR) was performed for p-value adjustment. Labels of statistically significant changed features were color-annotated, red for increasing and green for decreasing with q-value asterisks. *P &lt; 0.05, **P &lt; 0.01, ***P &lt; 0.001. Figure S8 Behavior deviation for male flies from female flies. A, Effect size for locomotion behavior deviation for w1118 male flies compared with w1118 female flies. B, Effect size for social behavior deviation for w1118 male flies compared with w1118 female flies. C, Effect size for locomotion behavior deviation for OR male flies compared with OR female flies. D, Effect size for social behavior deviation for OR male flies compared with OR female flies. E, Effect size for locomotion behavior deviation for CS male flies compared with CS female flies. F, Effect size for social behavior deviation for CS male flies compared with CS female flies. Statistical analysis: A modified Cohen’s d was used to calculate the effect size of behavioral deviation for male flies compared female flies from 3 experimental replicates. ANOVA were used to identify statistical significantly changed behavior metrics given experimental replicate, fly gender. False discovery rate (Benjamini/Hochberg FDR) was performed for p-value adjustment. Labels of statistically significant changed features were color-annotated, red for increasing and green for decreasing with q-value asterisks. *P &lt; 0.05, **P &lt; 0.01, ***P &lt; 0.001.