Fig. 4

In vitro pharmacological effect. The HepG2 cells were treated with various BBR formulations at an equivalent BBR concentration of 1 μg mL⁻¹ for 8 h at 37 °C in 5% CO₂. a The induction of p-AMPK (pink), InsR (red) and LDLR (green) protein expression by different BBR formulations was probed simultaneously with anti-InsR, LDLR and p-AMPK antibodies and visualized with C2t Nikon fluorescent microscope (Morrell, USA). b The mRNA expression of AMPK, InsR, AKT, and LDLR genes was evaluated by RT-PCR. The results were normalized to GAPDH. c The protein expression of AMPK, p-AMPK, InsR, p-InsR, AKT, p-AKT, and LDLR was tested using western blot analysis. The results were normalized to GAPDH as density ratio. Data are presented as mean ± SEM (n = 6), *p < 0.05, **p < 0.01, ***p < 0.001, vs NC group, p-values were calculated by unpaired two-sided Student’s t-test. Scale bars, 100 μm (a)