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Electromagnetic interference (EMI) and radiation of electronic devices are ubiquitous, which are potentially hazardous to the normal operation of electronic equipment and human health. MXenes are extremely attractive in the preparation of EMI shielding materials due to their excellent metallic conductivity and tunable surface chemistry. Herein, by virtue of the designed nanostructure and regulation of interface interactions, we fabricated flexible Fe3O4@Ti3C2Tx MXene/3,4-dihydroxyphenylacetic acid (DOPAC)-epoxidized natural rubber (ENR) elastomers (FMDE) with 3D segregated interconnected structures. The elaborately designed metal-ligand coordination crosslinking between Fe3O4 nanoparticles and DOPAC ligand molecules provides strong interfacial interactions, resulting in significantly reinforced mechanical properties. Compared with Ti3C2Tx/ENR elastomers, the maximum tensile strength and toughness of FMDE are elevadted by ~306% and 475%, respectively. Moreover, the 3D segregated conductive network constructed by Fe3O4@Ti3C2Tx nanoflakes resulted from volume exclusion effect of ENR latex and the introduction of magnetic Fe3O4 nanoparticles with enhanced electromagnetic wave absorption greatly improved the EMI shielding performance of FMDE, exhibiting an excellent EMI shielding effectiveness of up to 58 dB in the X band (8.2–12.4 GHz) and stable EMI shielding capability during repeated deformations. This work provides a promising strategy for the design and manufacture of novel flexible EMI shielding materials.