Fig. 1

Study design. A Classification of experimental groups. In the negative and positive control groups, sperm were incubated in HTF + 0.4% BSA for 30 min. After 30 min, sperm in DMSO/CAG-GFP and DMSO/gRNA-Cas9 groups were treated with 3% DMSO and 20 ng/µl plasmid for 15 min at 4 °C. In the sham groups, sperm were incubated in a c-TYH medium with different concentrations of MBCD (0, 0.75, 1, and 2 mM) for 30 min. In the treatment groups, sperm were incubated in a c-TYH medium with different concentrations of MBCD (0.75, 1, and 2 mM) in the presence of 20 ng/µl plasmids for 30 min. B Overview of assessment performed on experimental groups. C Details of the sperm incubation medium for sham and treatment groups. D Schematic view of DMSO treatment of sperm. EC: extracellular; TR: transfection rate