Figure 5.

Corylin Inhibits de novo Lipids Synthesis by Regulating SREBPs Activity. (A) HL-7702 cells were treated with HSP90 inhibitors with different structure backbone. Only corylin did not cause cytotoxicity. (B) HL7702 cells were depleted of sterols by incubating in medium D for 24 h, and then switched to medium D containing indicated concentration of corylin for 4 h, or corylin (3 μg/ml) for different time, the whole cell extracts underwent immunoblotting with indicated antibodies. (C-D) HL7702 cells were depleted of sterols by incubating in medium D for 24 h, and then switched to medium D containing 3 µg/ml corylin for 4 h. The expression of various genes was analyzed by qRT-PCR. (E) HL7702 cells were depleted of sterols by incubating in medium D for 24 h, and then switched to medium D containing 3 μg/ml corylin or 2 μM lovastatin for 16 h. Acetic acid sodium salt 1-¹⁴C was directly added into the medium and incubated for additional 2 h. (F) 293T cells were transfected with Myc-HSP90β and HA-Akt for 24 h, after the treatment, the cells were incubated with medium D containing corylin for another 4 h. Cells were lysed and pulled down by Myc antibody. HL-7702 hepatocytes were treated with GSK3β inhibitor SB216763 (10 µM) (G) or CHIR-9902 (10 µM) (H) for 1 h, the cells were switched to medium D supplemented with inhibitors plus vehicle, or 3 µg/ml corylin for 4 h. The whole cell extracts underwent immunoblotting with indicated antibodies. (I) HL-7702 hepatocytes were transfected with GSK3β siRNA for 48 h, after the treatment, the cells were switched to medium D treated with 3 µg/ml corylin for 4 h. (J) HL-7702 hepatocytes were transfected with HA-Akt for 24 h, after the treatment, the cells were switched to medium D treated with 3 µg/ml corylin for 4 h. Whole cell extracts underwent immunoblotting with indicated antibodies. Error bars are represented as mean ± SEM. Statistical analyses were done with one-way ANOVA (Dunnett's post test) (A and C). *p < 0.05, **p < 0.01, ***p < 0.001 vs DMSO.