Fig. 3

Disruption of GCN5L1 altered the activity of the ROS–ERK–FOXO1 axis in MLE-12 cells. A Q-PCR detection of Foxo1 mRNA expression in WT, GCN5L1-KO MLE-12 cells. B Representative immunoblot image of Foxo1 protein expression levels in WT, GCN5L1-KO MLE-12 cells and M2 cells infected with lentivirus expressing exogenous GCN5L1 (M2 + GCN5L1). C Representative immunoblot images of ERK and p-ERK protein expression levels in WT, GCN5L1-KO MLE-12 cells and M2 cells infected with lentivirus expressing exogenous GCN5L1 (M2 + GCN5L1). D Cellular ROS levels of WT, GCN5L1-KO MLE-12 cells, and M2 cells infected with lentivirus expressing exogenous GCN5L1 (M2 + GCN5L1). Cells were incubated with CM-H2DCFDA and measured by FACS; mean fluorescence intensity was normalized to WT group. E Representative immunoblot images of ERK, p-ERK, Foxo1, and Cebpα protein expression levels in WT cells, M2 cells treated with DPI (M2 + DPI) or SCH772984 (M2 + SCH772984), and M2 cells infected with lentivirus expressing exogenous Foxo1 (M2 + Foxo1). F Q-PCR assays of the RNA levels of surfactant-related genes in WT cells, M2 cells treated with DPI (M2 + DPI) or SCH772984 (M2 + SCH772984), and M2 cells infected with lentivirus expressing exogenous Foxo1 (M2 + Foxo1). The results are expressed as the mean ± SD of three independent experiments; ns, not significant; *P < 0.05,**P < 0.01; t-test