Figure 5

Similarities and differences in the effects of imeglimin and metformin on gene expression in HepG2 cells. (A–C) HepG2 cells were stimulated with 0.25 or 3 mmol/l metformin or imeglimin, with 1 mmol/l AICAR, or with vehicle for 12 h, after which RNA was isolated from the cells and subjected to RNA-seq analysis (n = 3 independent replicates). Multidimensional scaling analysis (A) and hierarchical clustering analysis (B) of the sequencing data, as well as a logarithmic scatter plot of the fold changes in gene expression (TPM > 10) induced by metformin or imeglimin at 3 mmol/l relative to the vehicle control (C), are shown. (D) RT-qPCR analysis of expression of the four genes indicated in (C) for cells treated with 3 mmol/l metformin or imeglimin, with 1 mmol/l AICAR, or with vehicle. (E) RT-qPCR analysis of expression of the four genes indicated in (C) for cells transfected with LKB1-targeting or control siRNAs for 72 h and then stimulated with vehicle or with 3 mmol/l metformin or imeglimin for 12 h. All RT-qPCR data are means ± SEM (n = 3 independent replicates). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns (one-way ANOVA followed by Tukey’s test).