Figure 3.

Interactive effects of KB and citric acid cycle-related metabolites on L6 cells differentiation. Cells were cultured for 4 d in differentiation medium supplemented with or without KB (1 mM LA and 4 mM HB), PA (16 mM pyruvate), αK (16 mM α-ketoglutarate), and OA (16 mM oxaloacetate). (A) Metabolic pathways of KB (acetoacetate and β-hydroxybutyrate) and intermediate metabolites of the citric acid cycle. (B) Relative mRNA expression levels of differentiation markers (Lmod2 and Ckm) were quantified using RT-qPCR. (C) Cell viability was determined using neutral red assay. Values are expressed as the mean ± SD of three independent experiments. Data were analyzed using one-way ANOVA followed by Tukey–Kramer multiple comparison test. Different letters above the bars indicate statistically significant differences at the 5% level.