Purpose: To uncover the mechanism of 4-nonylphenol-induced impairment of sertoli cells, we used RNA sequencing technology to compare the small RNA expression differences between the control groups and the 4-nonylphenol treated groups. Sertoli cells small RNA profiles of the control groups and the 4-nonylphenol treated groups were generated by deep sequencing, in triplicate. The library quality was determined using a Bioanalyzer 2100 (Agilent). The Illumina BGISEQ platform was used for small RNA sequencing. The quality of RNA-seq reads was examined using FastQC. Overall design: 4-nonylphenol-treated and control sertoli cells small RNA profiles of 24h were generated by deep sequencing, in triplicate.