Predation on anadromous salmon can have important consequences for both predators and prey. Salmon provide large seasonal pulses of energy and nutrients via carcasses, eggs and juveniles to many freshwater consumers, and conversely, predation can represent a significant source of mortality for juvenile salmon. Recent declines of Chinook salmon (Oncorhynchus tshawytscha) populations in Alaska have raised concern that predation might inhibit their recovery. Here, we quantify patterns of predation by freshwater fishes on juvenile salmon across seasons, habitats, predator sizes and streamflow levels in the Arctic-Yukon-Kuskokwim region of Alaska. We analysed piscivore stomach contents and identified prey using DNA sequence “barcoding.” In coastal rivers, juvenile pink (O. gorbuscha) and chum (O. keta) salmon contributed heavily to Arctic grayling (Thymallus arcticus) and Dolly Varden char (Salvelinus malma) diets, coho salmon (O. kisutch) prey were rare, and Chinook salmon were not detected. In interior rivers, Arctic grayling, burbot (Lota lota) and northern pike (Esox lucius) consumed small numbers of Chinook salmon. Predation on Chinook salmon was documented disproportionately in sloughs during a summer of exceptionally high streamflow. Dietary and distributional patterns suggested northern pike and burbot may exclude salmon from sloughs in low-gradient river reaches that would otherwise provide suitable rearing habitat. The data also provided tentative support for the hypothesis that high streamflow induces juvenile Chinook salmon to move from mainstem habitats into sloughs, where they face an increased risk of mortality. Incorporating predation risk into climate adaptation, fisheries management and habitat restoration decisions may help to facilitate Chinook salmon recovery.