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For tissues, we collected fetal tissues from four healthy pregnant Holstein cows at a slaughterhouse. After the slaughter, we extracted fetal tissues to examine their normal development and measured the crown-rump length (CRL) to estimate gestational age. The gender was identified based on the position of the gonads within the abdominal cavity, anatomical structures, and relationships with adjacent organs (mesonephros and/or kidneys). Two male and two female fetuses were identified (13-16 cm CRL), nine different tissues were collected, including the forebrain, hindbrain, heart, liver, lungs, skeletal muscle (hind limb), kidneys, rumen, and testes. Cells were collected by tryple-select enzyme and the lyophilized cells were sent for sequencing. Transcriptome (RNA) sequencing was performed by Tianjin Nuohe Zhiyuan Biotechnology Co., LTD, Tianjin, China. Sequencing libraries were generated using NEBNext UltraTM RNA Library Prep Kit for Illumina (NEB, USA) following manufacturer’s recommendations and index codes were added to attribute sequences to each samp |
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