The ability to gate (i.e., allow or block) droplet and fluid transport in a directional manner represents an important form of liquid manipulation and has tremendous application potential in fields involving intelligent liquid management. Inspired by passive transport across cell membranes which regulate permeability by transmembrane hydrophilic/hydrophobic interactions, macroscopic hydrophilic/hydrophobic Janus-type membranes are prepared by facile vapor diffusion or plasma treatments for liquid gating. The resultant Janus membrane shows directional water droplet gating behavior in air-water systems. Furthermore, membrane-based directional gating of continuous water flow is demonstrated for the first time, enabling Janus membranes to act as facile fluid diodes for one-way flow regulation. Additionally, in oil-water systems, the Janus membranes show directional gating of droplets with integrated selectivity for either oil or water. The above remarkable gating properties of the Janus membranes could bring about novel applications in fluid rectifying, microchemical reaction manipulation, advanced separation, biomedical materials and smart textiles.