This data release presents chemical and biological results from an investigation of the fish uptake and effects from exposure to per- and polyfluoroalkyl substances (PFAS) from groundwater contaminated by fire training activities on Cape Cod, Massachusetts. Twenty-one day exposure experiments were conducted from July to August 2019 and from August to September 2021 using groundwater from an uncontaminated reference site (REF) and PFAS contaminated sites near a legacy fire training area (FTA-1, FTA-2). Details for the groundwater sampling sites and well construction are presented in Table 1. During the course of the experiments, PFAS mixture concentrations and composition in groundwater and in fish plasma were measured using 2 separate analytical methods (water and plasma) and additional contaminants of concern (COC) also were measured (Table 2). PFAS quality assurance data for water analysis are presented in Table 3, and PFAS results for water samples from both experiments are presented in Table 4. Water results for COC are presented in Table 5. Fish biomarker results from 2019 and 2021 are presented in Table 6. Fish plasma PFAS results from 2021 are presented in Table 7. PFAS measured in 2021 fish plasma are listed in Table 8. Comments on 2021 fish plasma analysis are listed in Table 9. Quality assurance data for 2021 fish plasma PFAS analysis are presented in Tables 10-14.