ABSTRACT In the operation of the proton exchange membrane fuel cell (PEMFC) with dead-ended anode (DEA), the cathode relative humidity affects the accumulation state of water and nitrogen on the anode side, which in turn seriously affects the cell’s working performance. In order to investigate the mechanism of the cathode relative humidity on the PEMFC performance, numerical simulation and segmented in-situ measurement methods were used in this study. The results of the study showed that the lower cathode relative humidity causes insufficient water content in the PEMFC. This can further lead to localized dehydration of the membrane, which causes degradation of the DEA-PEMFC performance. Moreover, the higher cathode relative humidity increases the water content on the cathode side of the PEMFC. This not only increases the membrane permeability to nitrogen but also accelerates the water concentration back-diffusion process. The water and nitrogen that penetrate into the anode side can occupy the gas transport channel and hinder hydrogen transport, resulting in the insufficient localized gas supply in the DEA-PEMFC. This finally results in lower transient voltages and significantly shorter stabilized operation times. Therefore, reasonable control of cathode relative humidity is required to ensure long-term efficient and stable operation of the DEA-PEMFC.