Fig. 8.

GCN5L1 knockout resulted in excessive ROS signaling in the suppression of gluconeogenesis. a Cellular ROS levels were measured by FACS. Primary hepatocytes were incubated with vehicle (Veh), Diphenyleneiodonium (DPI) or MitoTEMPO (Tempo) for 30 min before incubated with CM-H2DCFDA. Mean fluorescence intensity was normalized to Con group with Veh treated (n = 3 independent experiments). b Mitochondrial hydrogen peroxide levels were measured using isolated mitochondria from DPI or MitoTEMPO treated primary hepatocytes (n = 3 independent experiments). c Primary hepatocytes were treated with DPI or MitoTEMPO for 30 min and cell lysates were analyzed by immunoblotting of p-ERK (n = 3 independent experiments). d, e Primary hepatocytes were incubated with Veh or MitoTEMPO overnight and followed by another 4 h incubation in glucose production medium for RNA analysis (d) and glucose analysis (e) (n = 3 independent experiments). f Cellular ROS levels were measured in hepatocytes at 36 h after adenovirus infection. Mean fluorescence intensity was normalized to Con group with vector adenovirus infected (n = 3 independent experiments). g Mitochondrial hydrogen peroxide levels were measured using isolated mitochondria from adenovirus infected primary hepatocytes (n = 3 independent experiments). h Primary hepatocytes were isolated and transfected with adenoviral expression of control, GCN5L1 (G) or Mt-GCN5L1 (MTG), glucose production was analyzed at 36 h after transfection (n = 3 of independent experiments). Values are expressed as mean ± s.e.m. *P < 0.05, **P < 0.01 by Student’s t-test