Fig. 4: MiR-98 could inhibit the microglial phagocytosis of neuron after OGD/R in vitro.

a The procedure of primary microglia transfection, OGD/R and physical contact co-cultured with neuron. b The target sequence of hsa-miR-98 and WT & Mut PAFR 3′-UTR. c The effect of hsa-miR-98 on PAFR expression via dual-luciferase report system. d The transfection efficiency of negative control (NC-FITC). e The transfection efficiency of miR-98 agomir. f–g The transfection efficiency of PAFR-siRNA. h–i The CellTracker^TM Green-labeled microglial phagocytosis of pHrodo^TM Red Dextran dye after treated with OGD 3 h and reoxygenation during the time course from 30 min to 150 min and quantification of the phagocytic %area. (Scale Bar = 100 μm). j Representative immunological staining of NeuN and MAP2 after primary neuron treated with 2 h OGD and reoxygenation for 24 h. (Scale Bar = 100 μm). k The representative images of the effects of overexpressing miR-98 and downregulating PAFR on the microglial phagocytosis of neurons after treated with OGD 2 h and reoxygenation for 24 h. The white arrows mean representative microglial phagocytosis of neurons. (Scale Bar = 50 μm). l–m The representative phagocytes of microglia and quantification of the percentage of phagocytes/total microglia numbers. (Scale Bar = 50 μm) Data are shown as means±S.E.M., ^*P, ^&P < 0.05; ^**P < 0.01; ^***P, ^###P, ^$$$P < 0.001 vs corresponding control. (n = 3 independent experiments).