In the kidney, the sodium-glucose cotransporters SGLT2 and SGLT1 are thought to account for >90% and ~3% of fractional glucose reabsorption (FGR), respectively. Yet, euglycemic humans treated with an SGLT2 inhibitor maintain an FGR of 40-50%, mimicking values in Sglt2 knockout mice. Here we show that oral gavage with a selective SGLT2 inhibitor (SGLT2-I) dose-dependently increased urinary glucose excretion (UGE) in wild-type (WT) mice. The dose-response curve was shifted leftward and the maximum response doubled in Sglt1 knockout (Sglt1-/-) mice. Treatment in diet with the SGLT2-I for 3 weeks maintained 1.5-2 fold higher urine glucose/creatinine ratios in Sglt1-/- vs. WT mice, associated with a temporarily greater reduction in blood glucose in Sglt1-/- vs. WT after 24h (-33 vs -11%). Subsequent inulin clearance studies under anesthesia revealed free plasma concentrations of the SGLT2-I (corresponding to early proximal concentration) close to the reported IC50 for SGLT2 in mice, which were associated with FGR of 64±2% in WT and 17±2% in Sglt1-/-. Additional i.p. application of the SGLT2-I (maximum effective dose in metabolic cages) increased free plasma concentrations ~10 fold and reduced FGR to 44±3% in WT and to -1±3% in Sglt1-/-. Absence of renal glucose reabsorption was confirmed in male and female Sglt1/Sglt2 double knockout mice. In conclusion, SGLT2 and SGLT1 account for renal glucose reabsorption in euglycemia, with 97% and 3% being reabsorbed by SGLT2 and SGLT1, respectively. When SGLT2 is fully inhibited by the SGLT2-I, increase in SGLT1-mediated glucose reabsorption explains why only 50-60% of filtered glucose is excreted.