Anti angptl3

Cells were treated with vehicle or tangeretin (20 and 40 μM) for 24 h. Total cellular proteins were extracted using RIPA buffer (Thermo Fisher Scientific, Rockford, IL, USA). For nuclear extract preparation, cells were harvested, and nuclear proteins were prepared using NE-PER nuclear and cytoplasmic extraction reagent (Thermo Fisher Scientific, Rockford, IL, USA). The proteins were separated by 10% or 12% SDS–PAGE and transferred onto a PVDF membrane (PerkinElmer, Boston, MA, USA), followed by incubation with specific primary antibodies for human proteins, including anti-ANGPTL3 (ABclonal, Woburn, MA, USA), anti-LXRα (Abcam, Cambridge, MA, USA), anti-HNF-1α (Cell Signaling Technology, Danvers, MA, USA), anti-HDAC2 (GeneTex, Irvine, CA, USA), and anti-actin (Millipore Sigma, St. Louis, MO, USA). The blots were incubated with the appropriate horseradish peroxidase (HRP)-conjugated secondary antibodies (GeneTex, Irvine, CA, USA) at room temperature. The protein signals were detected using Amersham ECL^TM prime Western reagents (GE Healthcare, Buckinghamshire, UK), and chemiluminescence-exposed Amersham Hyperfilm^TM ECL film (GE Healthcare, Buckinghamshire, UK) was analyzed.

Cells were harvested, and total cellular protein was extracted using RIPA buffer (Thermo Fisher Scientific). Nuclear proteins were prepared using a Nuclear Extract Kit (Active Motif, Carlsbad, CA, USA). The proteins from each sample were separated via 10% SDS–PAGE and transferred to PVDF membranes (Cytiva). The membranes were incubated with the following specific primary antibodies: anti-ANGPTL3 (Cat#A5225, RRID:AB_2863493, for the human cell line samples) (ABclonal, Woburn, MA, USA), anti-ANGPTL3 (Cat#PA5-72812, RRID:AB_2718666, for the zebrafish samples) (Thermo Fisher Scientific), anti-LXRα (Cat#ab41902, RRID:AB_776094) (Abcam, Cambridge, UK), anti-HDAC2 (Cat#GTX112957, RRID:AB_1950480) (GeneTex, Irvine, CA, USA), anti-Actin (Cat#MAB1501, RRID:AB_2223041) (Millipore Sigma-Aldrich) and anti-HNF-1α (Cat#89670, RRID:AB_2728751), anti-β-actin (Cat#8457, RRID:AB_10950489) (Cell Signaling Technology, Danvers, MA, USA). After that, the membranes were incubated with the appropriate horseradish peroxidase (HRP)-conjugated goat anti-rabbit (Cat#GTX213110-01, RRID:AB_10618573) (GeneTex) or anti-mouse (Cat#7076, RRID:AB_330924) (Cell Signaling Technology) IgG secondary antibodies. The membranes were rinsed with Amersham ECL^TM Prime Western blotting detection reagent, and the chemiluminescent signal bands were visualized with Amersham Hyperfilm^TM ECL (Cytiva).