MARCH5 regulation of autophagy through AKT-FOXO3 signaling. (A) Upregulation of the genes involved in the AKT pathway in the MARCH5 KD group vs the scramble group in the pathway analysis of the RNA-seq sample. (B) Upregulation of AKT and p-Akt, along with a decrease in the FOXO3 level in the MARCH5 KD groups. (C) Inhibition of p-AKT upon treatment of cells with increasing concentrations of MK-2206 for 24 hours leads to a dose-dependent increase in FOXO3 and LC3I levels. (D) Protein interaction of MARCH5 with FOXO3 as demonstrated by immunoprecipitation. (E) FOXO3 interaction with HA-tag vector targeting MARCH5 as demonstrated by coimmunoprecipitation. (F) Decreased MARCH5, SMAD2, and ATG5 expression following FOXO3 KD. Data are shown as mean ± standard error of the mean of 3 independent experiments. ∗P < .05; ∗∗P < .01; ∗∗∗P < .001.