The effect of 2HG level on transcriptome at different stages of male mouse meiosis

We found that L2-hydroxyglutarate (2HG) level is 15 times higher in pachytene-diplotene (PD) stage as compared to leptotene-zygotene (LZ) stage. Since 2HG shows epigenetic effects we asked if the addition of 2HG to LZ cells or the decrease of 2HG level in PD cells will cause changes in gene expression. Overall design: LZ and PD cells were isolated using flow cytometry from mice expressing tdTomato in germ cells. LZ cells were cultured for 6 days without and with 0.3 mM cell-permeable L2HG. PD cells were cultured for 48 hours without and with 24 mM oxamate, which reduces 2HG level to that of LZ cells. Three independant experiments were performed. RNA was isolated using Qiagen kit and RNA seq was performed .