Fig. 6. The effect of GC7 is fully reversible.

Confluent PCT cells were treated or not with 30 µM GC7 for 24 h and then GC7 was removed and cells were maintained for an additional period of 72 h and analysed. a, b Cells were analysed using Seahorse technology to evaluate oxygen consumption (a) and extracellular acidification (b) in the presence of 10 mM glucose. Addition of other compounds are indicated. Dots displayed mean ± SD of three independent experiments. *p < 0.05 evaluated using Mann–Whitney. c At the end of treatment cells were deprived in glucose and the efflux of glucose was measured in the next 8 h. d Measurement of cell glucose consumption corresponding to the difference between measurement of the media glucose content decrease (in the presence of glucose in media) and media glucose content increase (after glucose deprivation). Dots displayed mean ± SD of three independent experiments. *p < 0.05 evaluated using Mann–Whitney tests. “Ctrl rev” are control cells at 72 h without GC7 treatment and “GC7 rev” correspond to cells 72 h after GC7 removal. “Ctrl” and “GC7” are cells analysed after 24 h. e Western blot analysis of GLUT1 in membrane-enriched protein lysate from PCT cells following a 24 h GC7 treatment (30 µM) and after 72 h of GC7 washout (72 h rev). β-actin was used as a loading control. MW molecular weight. *p < 0.05 evaluated using Mann–Whitney test.