Figure 4.

Transcriptome analysis of DLE in vitro revealed that DLE affected gluconeogenesis via the AKT/FOXO1 signaling pathway. (A) Hierarchical clustering, (C) PCA, and (D) tSNE analysis of all groups, using the normalized read counts of RNA-mediated oligonucleotide annealing, selection, and ligation with next-generation sequencing that corresponded to all DEG unions as the input. (B) The volcano plots of DEGs induced by the indicated concentrations of DLE (upregulation in red and downregulation in green). (E) Line chart and heatmap of dose-dependent DLE regulated genes (upregulated in red and downregulated in blue). (F) Enriched KEGG pathways in DLE regulated dose responsive genes. (G) RT-qPCR results of 200 μg/mL DLE-regulated FOXO1-related genes (n = 3). (H) Western blot analysis of the protein expression and phosphorylation levels of FOXO1 (n = 3), AKT (n = 4), and GSK3β (n = 3). The results were quantified by ImageJ software, the data were quantified by ImageJ. (I) The inhibitory effects of DLE on the glucose production in HepG2 cells treated with 200 μg/mL DLE for 24 h (n = 3). The data are shown as the mean ± SD. Statistical analyses were conducted using paired Student's t-tests. ∗P < 0.05, ∗∗P < 0.01 vs. the DMSO-treated group. PKB/AKT, protein kinase B; FOXO1, forkhead box O1; GSK3β, glycogen synthase kinase 3 β; DMSO, dimethyl sulfoxide.