Nitrogen (N) and phosphorus (P) commonly stimulate phytoplankton production in lakes, but recent observations from lakes from an agricultural region suggest that nitrate may have a subsidy-stress effect on chlorophyll a (Chl a). It is unclear, however, how generalizable this effect might be. Here, we analyzed a large water quality dataset of 2385 lakes spanning 60 regions across 17 states in the Northeastern and Midwestern U.S. to determine if N subsidy-stress effects on phytoplankton are common and to identify regional landscape characteristics promoting N stress effects in lakes. We used a Bayesian hierarchical modeling framework to test our hypothesis that Chl a–total N (TN) threshold relationships would be common across the central agricultural region of the U.S. (“the Corn Belt”), where lake N and P concentrations are high. Data aggregated across all regions indicated that high TN concentrations had a negative effect on Chl a in lakes with concurrent high total P. This large-scale pattern was driven by relationships within only a subset of regions, however. Eight regions were identified as having Chl a–TN threshold relationships, but only two of these regions located within the Corn Belt clearly demonstrated this subsidy-stress relationship. N stress effects were not consistent across other intense agricultural regions, as we hypothesized. These findings suggest that interactions among regional land use and land cover, climate, and hydrogeology may be important in determining the synergistic conditions leading to N subsidy-stress effects on lake phytoplankton.