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Figure S1 Effects of bilobetin on TG dynamics in rats. The uptake of Intralipid-TG by RQ (A) and mWAT (B) in overnight-fasted (basal) or euglycaemic–hyperinsulinaemic clamp (clamp) rats. Normal, standard diet; HFD, high-fat diet; BIL14, HFD + 14 day bilobetin treatment; BIL4, HFD + 4 day bilobetin treatment; BIL4M, BIL4 rats treated with PPARα inhibitor (MK886); BIL4H, BIL4 rats treated with PKA inhibitor (H89). Data are means ± SE. n = 6. aP < 0.01 versus HFD group (ANOVA).

Figure S2 Effects of bilobetin on rates of uptake (A) and storage (B) of NEFA by mWAT and RQ in rats. Normal, standard diet; HFD, high-fat diet; BIL14, HFD + 14 day bilobetin treatment; BIL4, HFD + 4 day bilobetin treatment; BIL4M, BIL4 rats treated with PPARα inhibitor (MK886); BIL4H, BIL4 rats treated with PKA inhibitor (H89). Data are means ± SE. n = 6. aP < 0.01 versus HFD group (ANOVA).

Figure S3 Effects of bilobetin on expression of PPARα in RQ (A), PPARγ in mWAT (B) and RXRα in liver (C). Relative expression ratios are expressed as fold changes of protein content compared with normal group. Normal, standard diet; HFD, high-fat diet; BIL14, HFD + 14 day bilobetin treatment; BIL4, HFD + 4 day bilobetin treatment; BIL4M, BIL4 rats treated with PPARα inhibitor (MK886); BIL4H, BIL4 rats treated with PKA inhibitor (H89). Data are means ± SE. n = 6.

Figure S4 (A) Competitive binding curve of bilobetin (blue) and fenofibrate acid (purple) to PPARα ligand binding domain. Values are expressed as the 520 nm/495 nm emission ratio at a series of concentrations (1 nmol·L−1–1 mmol·L−1) of bilobetin. (B) Competitive binding curve of bilobetin (blue) and pioglitazone (purple) to PPARγ ligand binding domain. Values are expressed as the polarization values (mP) at a series of concentrations (1 nmol·L−1–1 mmol·L−1) of bilobetin. Values are means ± SE. n = 3.

Figure S5 Effects of bilobetin on cAMP level, PKA activity and AC activity. Cells were incubated with bilobetin (1 μmol·L−1) or forskolin (1 μmol·L−1) or forskolin plus bilobetin (each 1 μmol·L−1) for 0, 0.5, 1, 4, 12 and 24 h, and then the intracellular cAMP level (A) and PKA activity (B) were determined in cytoplasm. Data are means ± SE, n = 6. aP < 0.01 versus 0 h of corresponding group; bP < 0.01 versus corresponding time point of forskolin group (ANOVA). In another experiment, hepatocytes were incubated with forskolin (1 μmol·L−1) plus bilobetin (0, 10−7, 10−6, 10−5, 10−4 mol·L−1) for 1 h, and then the changes of cAMP level (C) and PKA activity (D) in cytoplasm were measured. Data are means ± SE, n = 6. aP < 0.01 versus forskolin (1 μmol·L−1)-treated cells. (E) AC activity. Data are means ± SE, n = 6. aP < 0.01 versus 10−6 mol·L−1 bilobetin-treated cells.

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