The stromal interaction molecules STIM1 and STIM2 sense a decreasing Ca 2+ concentration in the lumen of the endoplasmic reticulum and activate Ca 2+ channels in the plasma membrane. In addition, at least 2 reports suggested that STIM1 may also interact with the inositol 1,4,5-trisphosphate (IP 3 ) receptor. Using embryonic fibroblasts from Stim1 −/− , Stim2 −/− and wild-type mice, we now tested the hypothesis that STIM1 and STIM2 would also regulate the IP 3 receptor. We investigated whether STIM1 or STIM2 would be the luminal Ca 2+ sensor that controls the loading dependence of the IP 3 -induced Ca 2+ release. Partial emptying of the stores in plasma-membrane permeabilized cells resulted in an increased EC 50 and a decreased Hill coefficient for IP 3 -induced Ca 2+ release. This effect occurred both in the presence and absence of STIM proteins, indicating that these proteins were not the luminal Ca 2+ sensor for the IP 3 receptor. Although Stim1 −/− cells displayed a normal IP 3 -receptor function, agonist-induced Ca 2+ release was reduced. This finding suggests that the presence of STIM1 is required for proper agonist-induced Ca 2+ signaling. Our data do not provide experimental evidence for the suggestion that STIM proteins would directly control the function of the IP 3 receptor.