Fig. 4: Enhanced renal Sglt2 expression and glucose reabsorption in mice with global knockout of Nr1d1.

Female or male 16- to 18-week-old Bmal1^−/− mice and their WT (Bmal1^+/+) siblings were fed chow diet ad libitum. Renal cortex collected at the indicated times were used to measure mRNA (A: female, B: male; n = 3–4 animals per time point, Cosinar analysis for circadian changes) and protein (C). WB is representative of 3 independent gels. D Male 16- to 18-week-old Nr1d1^−/− and Nr1d1^+/+ siblings (n = 3/group) were fed chow diet ad libitum. Renal cortex collected at different times of day were used to measure protein levels. E BBMVs from male Nr1d1^+/+ and Nr1d1^−/− mice at 12:00 h were incubated with [¹⁴C]-αMG for 45 min to measure glucose uptake. Mean ± SD, n = 3 biological replicates/group, unpaired two-tailed t-test. RPTECs from male WT mice were treated with vehicle or hemin (5 µM) for 48 h and used to measure mRNA (F) or protein levels (G). Mean ± SD, n = 4 biological replicates/group, multiple unpaired t-test. Male 14- to 16-week-old Nr1d1^−/− and control mice were transduced intravenously with lentiviruses for expression of shBmal1 (2.5 × 10¹¹ GC/mouse) or shControl. After 2 weeks, renal cortices were used to measure mRNA (H) and protein (I) levels. Mean ± SD, n = 4 biological replicates per group, multiple unpaired t-tests. J BBMVs from shBmal1 transduced mice at 12:00 h as in (H, I) were incubated with 1 µCi/ml [¹⁴C]-αMG to measure glucose uptake. Mean ± SD, n = 4/group, unpaired two-tailed t-test. *p < 0.05, **p < 0.01 and ***p < 0.001; # p < 0.05, ## p < 0.01 and ### p < 0.001.