Fig. 7: Fasting induces Bmal1 expression.

Male 16- to 18-week-old Bmal1^−/− and Bmal1^+/+ mice were fed a high fat (HFD) diet ad libitum for 2 months (F0). They were then fasted at 12:00 h or at 24:00 h for 24 h (F24) and refed normal chow for 8 h after 24 h fasting. A Bmal1, Rev-erbα and Nrf1 in the nucleus, and Sglt2 in BBMV of the renal cortex were measured with western blotting (A up: gels at 12:00 h; down: gel at 24:00 h). B BBMVs prepared from mice subjected to a fasting and feeding regimen, at 12:00 (n = 5/6) or 24:00 (n = 4) h were incubated with [¹⁴C]-αMG for 120 min to measure glucose uptake. Mean ± SD, unpaired two-tailed t-test. C RPTECs isolated from Bmal1^−/− and Bmal1^+/+ mice were cultured in serum-containing medium (F0) or serum-free medium for 48 h (F48), then refed with 20% FBS serum for 8 h. The cells were used to measure mRNA levels. Values are mean ± SD, n = 4/group, unpaired two-tailed t-test. D RPTECs from male Bmal1^+/+ and Bmal1^−/− mice were cultured in serum-free medium for 48 h, then refed with 20% FBS containing medium for 8 h and used to measure protein with anti-Bmal1, anti-Rev-erbα, anti-Nrf1 (in the nucleus), anti-Sglt2 and anti-Gapdh (in the cell lysate) (up: gels; down: WB gel quantification of protein expression normalized to Gapdh). Mean ± SD, n = 4, unpaired two-tailed t-test. For all panels, *p < 0.05, **p < 0.01 and ***p < 0.001; #p < 0.05, ##p < 0.01 and ###p < 0.001.