Fig. 5

FG4592 inhibits the enzymatic activity of OGFOD1 and blocks its combination and hydroxylation of RPS23. A Molecular structure of FG4592. B and C Docking analyses of FG4592 covalent binding modes to PHD2 and OGFOD1. D The fluorescent staining images of OGFOD1 and different neural cell markers in healthy brains of mice or peri-infarct brains of mice after 24 hours of tMCAO. The NeuN and GFAP were markers of neurons and astrocytes, respectively. The brain slices of CX3CR1-GFP mice were used to exhibit the microglia. Scale bar, 100 μm, 40 ×. E The hydroxylation area of Pro-62 within RPS23 immunoprecipitated. Cells were dealt with vehicle or 100 μM FG4592 for 24 hours, and its split product was immunoprecipitated by anti-RPS23. F Representative western blots of RPS23 in the anti-OGFOD1 immunoprecipitates and input groups. Primary neuron cells underwent OGD for 3 hours and were treated with vehicle or FG4592 for 6 hours after reperfusion. The cell extracts were immunoprecipitated with OGFOD1 antibody or control IgG and immunoblotted for RPS23