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Immunofluorescence staining Primary murine chondrocytes and RAW 264.7 murine macrophage cell line were fixed with 3.7% PFA for 30 min at RT, permeabilized with TBS + 0.05% Tween 20 buffer for 30 min at RT, and blocked with 2.5% normal horse serum for 20 min at RT

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The DHE staining results also showed that FOXO4-DRI can effectively reduce the production of ROS in the vessels of aging mice (Figure 1F)