Data from : The Drosophila melanogaster Y-linked gene, WDY, is required for sperm to swim in the female reproductive tract. Yassi Hafezi, Arsen Omurzakov, Jolie A. Carlisle, Ian V. Caldas, Mariana F. Wolfner, Andrew G. Clark Department of Molecular Biology and Genetics, Cornell University, 227 Biotechnology Building, 526 Campus Rd., Ithaca, NY 14853-2703 Contact : YH at yh676@gmail.com or AGC at ac347@cornell.edu This dataset is shared under an Attribution 4.0 International (CC BY 4.0) license (https://creativecommons.org/licenses/by/4.0/). The material can be shared and built upon, but attribution to the original authors and a statement of changes made is required. Suggested citation : Hafezi, Y., Omurzakov, Arsen, Carlisle, J., Vasconcellos Caldas, I., Wolfner, M., & Clark, A. (2023). Data from: The Drosophila melanogaster Y-linked gene, WDY, is required for sperm to swim in the female reproductive tract [Dataset]. Cornell University Library eCommons Repository. https://doi.org/10.7298/VTR8-AB60 Related publication citation: Hafezi, Y., Omurzakov, A., Carlisle, J.A. et al. The Drosophila melanogaster Y-linked gene, WDY, is required for sperm to swim in the female reproductive tract. Commun Biol 7, 90 (2024). https://doi.org/10.1038/s42003-023-05717-x Changelog (see additional details below): Initial upload to eCommons: 2023-06-30 Additional files appended: 2023-09-21 Readme file updated with publication ciation: 2024-01-16 1. Sperm Distribution Images Image index and quantification are included (one .csv file). Images (.tiff files) of Protamine-GFP-labelled sperm from mutant and control males swimming within reproductive tracts of Canton-S females. Females were frozen after mating and reproductive tracts were dissected, fixed and stained. Images were captured by an Echo Revolve microscope. GFP fluorescent images are included for all samples; brightfield images of most PRY-related samples are also included. - PRY: experiments comparing three "PRY alleles" (G54.1, G55.8, and G54.7) and three "control alleles" (A1, A2, CanS) PRY image names format: slide_allele_sample_image - WDY: experiments comparing three "WDY alleles" (C104, F8, C3) and three "control alleles" (G107, Ytom, C7) WDY image names format: allele_slide_sample_image_channel - 2hASM: Sperm distribution in females 2 hours after start of mating - 24hASM: Sperm distribution in females 24 hours after start of mating - ONM: Sperm distribution in females after overnight mating - 30mASM: Sperm distribution in females 30 minutes after start of mating Files added to eCommons on 6/30/2023. 2. Sperm Head Swimming Videos Video index and quantifications are included (two .csv files). Videos (.mp4) of Protamine-GFP-labelled sperm from mutant and control males swimming within reproductive tracts of Canton-S females. Female reproductive tracts were dissected and live-imaged on an Echo Revolve microscope after females mated. The video files in this dataset contain no human voices; no transcripts are included. Video names format: allele_video Files added to eCommons on 6/30/2023 and 11/29/23. 3. Sperm Tail Beating Videos Includes .csv files (3 per experiment) outlining the batch structure of experiment, video index, and quantification data. Videos (.mov) of sperm dissected from male seminal vesicle or female uterus after mating and imaged under brightfield optics with an Olympus BX51WDI microscope and a 50x LMPLFLN objective. The video files in this dataset contain no human voices; no transcripts are included. - PRYvControl: experiment comparing three "PRY alleles" (G54.1, G55.8, and G54.7) and three "control alleles" (A1, A2, CanS). PRY image names format: slide_allele_sample_image - WDYvControl: experiment comparing three "WDY alleles" (C104, F8, C3) and three "control alleles" (G107, Ytom, C7) WDY image names format: allele_slide_sample_image_channel Files added to eCommons on 6/30/2023. 4. Sperm Transfer Images Image metadata (.txt file for each image set) and index with quantification (3 .csv files) are included. Images (.tif) of Protamine-GFP-labelled sperm within the reproductive tracts of Canton-S females. Female reproductive tracts were dissected, fixed and stained. Z-stacks were captured on a Leica DMRE confocal microscope. Image names format: Allele_Slide_Sample_Z-section_Channel Files added to eCommons on 6/30/2023. 5. WDY Codeml Analysis Files Files (.txt) used to run the positive selection analysis on WDY sequences in the codeml program of PAML 4.8 (Yang, 2007). PAML is a package of programs for phylogenetic analyses of DNA or protein sequences using maximum likelihood (see http://abacus.gene.ucl.ac.uk/software/paml.html). Sources of WDY sequences are listed in Supplementary Table S7 and sequences are included as a FASTA file (2023_09_21_WDY_nt_sequences.txt). "Other" folder includes additional output files generated by PAML. Files added to eCommons on 9/21/2023. 6. WDY Final Annotations PYMOL session (.pse) and protein database (.pdb) files indicating annotations of the conserved domains of D.melanogaster WDY based on the Alphafold protein structure prediction. Sequences of annotated domains are also included in Supplementary Table S6. Files added to eCommons on 9/21/2023.