Effect of OGT knockdown on gene expression in KGN ovarian granulosa cells

This study aimed to elucidate the spatiotemporal expression pattern of O-GlcNAc transferase (OGT) and its catalyzed O-GlcNAc modification in ovarian tissues, and to investigate their specific functions in ovarian granulosa cells. To this end, high-throughput RNA sequencing was performed to analyze genome-wide transcriptional changes following OGT knockdown in the human ovarian granulosa cell line KGN. Two specific siRNAs targeting OGT (siOGT1 and siOGT2) and a negative control siRNA (siNC) were designed. Each group included two biological replicates, resulting in a total of six transcriptomic libraries. Sequencing was carried out on the Illumina NovaSeq 6000 platform with paired-end 150-bp reads. By comparing the siOGT-treated groups with the siNC control group, 300 and 487 differentially expressed genes (DEGs) were identified in the siOGT1 and siOGT2 groups, respectively, with 176 DEGs overlapping between the two groups. Pathway enrichment analysis indicated that OGT knockdown may affect pathways related to immune defense response, cellular reaction to external stimuli, and cytokine-mediated signaling. This dataset provides a transcriptomic profile of ovarian granulosa cells under OGT interference and serves as a key resource for further investigation into the regulatory mechanisms of the OGT/O-GlcNAc axis in ovarian physiology and pathology. Overall design: To investigate the transcriptomic alterations following OGT knockdown, we transfected KGN ovarian granulosa cells with siOGT1 and siOGT2 separately, and harvested the samples for RNA-Seq analysis 48 hours post-transfection.