Uranium, a heavy metal and primordial radionuclide, is present in surface waters and soils both naturally and due to industrial activities. Uranium is known to be toxic to plants and its uptake and toxicity can be influenced by multiple factors such as pH and the presence of different ions. However, the precise role of the different ions in uranium uptake is not yet known. Here we investigated whether calcium influences uranium uptake and toxicity in the terrestrial plant Arabidopsis thaliana. To this end, A. thaliana plants were exposed to different calcium and uranium concentrations and furthermore, calcium channels were blocked using the calcium channel blocker lanthanum chloride (LaCl3). Fresh weight, relative growth rate, concentration of nutrients and uranium and gene expression of oxidative stress-related genes and calcium transporters were determined in roots and shoots. Calcium affected plant growth and oxidative stress in both control (no uranium) and uranium-exposed plants. In shoots, this was influenced by the total calcium concentration, but not by the different tested uranium concentrations. Uranium in turn did influence calcium uptake and distribution. Uranium-exposed plants grown in a medium with a higher calcium concentration showed an increase in gene expression of NADPH oxidases RBOHC and RBOHE and calcium transporter CAX7 after uranium exposure. In roots, these calcium-dependent responses in gene expression were not observed. This indicates that calcium indeed affects uranium toxicity, but only in shoots. In addition, a clear influence of uranium and LaCl3 (separately and combined) on the expression of calcium transporters was observed.