Investigation of Drosophila fruitless neurons that express Dpr/DIP cell adhesion molecules

Data tables of quantification of fru P1 ∩ dpr/DIP neurons. This is an excel data table with 10 sheets that contain: 1. Raw Data Brain: raw data and observation notes only for areas of the brain scored/counted. 2. Raw Data VNC: raw data and observation notes for areas of the VNC scored/ counted. 3. Raw Scoring Data: raw scoring observations of areas in both brain and VNC scored. 4. Scoring Data Processed: assigning numerical values between 0 and 1 to scoring observations. 5. Raw Count Data: raw cell counts for areas in both the brain and VNC. 6. Cell Count Calculations: math done to cell counts so they were between 0 and 1. 7. Full Data Set: both male and female image analysis data with scoring observations and cell counts as values between 0 and 1. 8. Male Only: image analysis results for males (used to create heatmaps). 9. Female only: image analysis results for females (used to create heatmap). 10. Heatmap_Behavior_Plot: supplemental figure of the heatmap and behavior plot where the count data and scoring data has been separated. This sheet also contains additional heat maps of the male and female image count data.

Data tables of quantification of behavioral phenotypes. This is an excel data table with five sheets that contain all the behavioral data as follows: 1. READ ME: This first sheet is to explain the following four tabs. Please note that TNTQA1, as opposed to TNTQ is used in the following sheets. They both refer to the inactive form of TNT, but TNTQA1 is our lab nomenclature. 2. TrpA1 male-female courtship behavior data. 3. TrpA1 male alone courtship behavior data. 4. TNT courtship behavior data. 5. TNT DAM activity locomoter behavior data: This tab contains line crossing reads from Drosophila Activity Monitors (DAM).

Data tables that include information from the single-cell RNA-seq study, including data matrices and UPSET analyses, in five sheets as follows: 1. Barcode rank plot and sequencing matrices. 2. Male 48 hr APF fru P1 neurons dpr/DIP gene-cell barcode matrix of log normalized and scaled gene counts. Columns are cell barcodes, rows are dpr/DIP genes. 3. Male 48 hr APF fru P1 neurons dpr/DIP expression matrix binarized by 0 or one where normalized, scaled expression >1 = 1 and expression <1 = 0. Data table is presented in transposed in format. Columns are dpr/DIP genes, rows are cell barcodes. 4. Numbers of single cells with increasing numbers of dprs/DIPs co-expressed, derived from data on sheet 3. 5. Upset plot of dpr/DIP expression combinations based on binarized expression matrix (sheet 3) where normalized, scaled expression >1 = 1 and expression <1 = 0. dpr/DIP expression dendrogram based on expression matrix presented in sheet 2.

Data tables of quantification of confocal data from sex hierarchy perturbations in fru P1 ∩ DIP-α neurons. This excel file has the quantification from the sex hierarchy perturbation analyses, with the following sheets: 1. Read Me: This first sheet is to explain the following tabs and to display the features scored. Representative confocal 3D projections are shown below. 2. Genotype and Condition: Description of genotypes, conditions, expected perturbations and changes. 3A. Scoring Summary of set one overexpressors: Overexpression of sex hierarchy related genes in ALL DIP-α cells plus a loss-of-function line (Control: DIP-α-GAL4; UAS>stop>GFP.Myr/+; fruFLP/+). 3B. The corresponding scoring raw data of 4A. 4A. Scoring Summary of set two overexpressors (with tubGal80): Overexpression of sex hierarchy related genes in DIP-α and fru P1 intersecting cells. (Control: DIP-α-GAL4; UAS>stop>GFP.Myr/+; fruFLP/tub>GAL80>). 4B. The corresponding scoring raw data of 5A (with tubGal80). 5. The conclusive tables of all scoring data and the corresponding statistics (p values from Fisher's Exact test; when p ≦ 0.05, the cell will be highlighted). 6. Confocal images showing staining controls: UAS-Fru^M constructs and expression pattern of Dip-alpha with UAS-GFP.

Data tables of quantification of confocal data of RNAi and dpr/DIP overexpression analyses, in four sheets as follows: 1. READ ME: This first sheet is to explain the following four tabs. 2. DIP-α- and DIPδ-GAL4 screen: This sheet contains all genotypes tested in the initial RNAi/overexpressor screen using both DIP-α and DIP-δ as the GAL4 driver. The controls are bolded. N = 5. This sheet contains the following columns: Genotype: Contains an abbreviated genotype, including the GAL4 driver and dpr/DIP knocked-down (RNAi) or overexpressed (UAS). Although not explicitly stated, all genotypes also contained a UAS > stop > GFP.Myr transgene as well as a fru^FLP transgene. Temperature Raised: The RNAi crosses were set up at 25°C where the flies were allowed to lay eggs for 2–3 days. Then, the parents were turned into a new vial, and the eggs were raised at 29°C. This was done to ensure that the RNAi was fully functional, as it was found to be less effective at 25°C. All overexpressor flies were raised at 25°C, as noted. Difficulty getting males: This was only an issue with the RNAi crosses at 29°C. Some of these crosses did not produce males at all, or produced far fewer males than females. Of note, these crosses always produced females with ease. Assayed at 16–24 hr: ‘Yes’ in this column indicated that five replicates per sex were dissected and stained as 16–24 hr adults. Assayed at 4–7 days: ‘Yes’ in this column indicated that five replicates per sex were dissected and stained as 4- to 7-day adults. Phenotype: Any phenotype observed when comparing the RNAi with its respective control. Notes: Relevant notes. 3.DIP-α-GAL4; DIP-ε-RNAi: Of all the genotypes tested, this was the only one to show a robust phenotype. The czi files were scored blind for conditions with and without tub>GAL80>. n = 20. A table summarizing the statistics is to the right of the raw data. See below for a more detailed description of how these phenotypes were scored. 4. UAS DIP-α screen: Although not explicitly stated, all genotypes also contained a UAS > stop > GFP.Myr transgene as well as a fru^FLP transgene. Each of the 7 GAL4s of interest were first crossed to UAS DIP-alpha (experimental) and Canton S (white) (control) with n = 5. The GAL4 stock was crossed to Canton S (white) (w;cs) as a control to eliminate balancers. The controls are bolded in this data sheet. 3 GAL4 drivers that showed potential phenotypes upon overexpressing DIP-alpha were identified for further study (DIP-β, DIP-δ, and DIP-ε). These genotypes were retested with an n = 15 and with tub>GAL80>. Upon further analysis, no robust phenotypes were observed and as a result, the images were not scored.