Fig. 6

TLR7/8 agonists inhibited ATP production via GSK3 α/β-hexokinase pathway. a, the ATP levels of sperm after culture in extender with 1 μmol/L R848 for a maximum of 60 min. b, the ATP levels of sperm in the upper layer and lower layer sperm with 1 μmol/L R848 for 30 min. c, the mitochondrial activity of sperm after culture in extender with 1 μmol/L R848 for a maximum of 60 min. d, the mitochondrial activity of sperm in the upper layer and lower layer sperm with 1 μmol/L R848 for 30 min. e, goat sperm were incubated with 1 μmol/L R848 for 0, 15, or 30 min, and 20 μg of protein lysates was analyzed for phospho-AKT, phospho-NFκB, phospho-GSK3α/β, total AKT, total NFκB, and total GSK3α/β expression by Western blotting. Tubulin was used as the loading control. Results are representative of three independent experiments. f, intensity of the phosphorylation of AKT, NFκB, and GSK3α/β in the upper layer and lower layer sperm induced by R848 for 30 min. g and h, columns represent the ratio of phosphorylated to total AKT, phosphorylated to total NFκB or phosphorylated to total GSK3α/β. Values are the mean ± SEM of three replicates. *P < 0.05 compared with the control