Ab61182

HepG2 cells were firstly cultured with GB1 or Wy14643 (HY-16995, MedChemExpress, Monmouth Junction, NJ, USA) in a petri dish (6 cm). Following PBS washing two times, the cells were lysed in lysis buffer to obtain total proteins. Equal amounts of protein were separated by electrophoresis and then transferred to polyvinylidene fluoride (PVDF) membrane (Millipore, Burlington, MA, USA). The membrane was subsequently blocked with 5% skim milk, followed by incubation with anti-PPARα (ab61182, Abcam, Shanghai, China), anti-SIRT6 (A3591, ABclonal, Woburn, MA, USA), anti-BCL-2 (T40056, Abmart, Berkeley Heights, NJ, USA), anti-β-actin (RM3002, Beijing Ray antibody biotech, Beijing, China), and anti-BCL-XL (T55050, Abmart, Berkeley Heights, NJ, USA) antibodies. Relative expression of the proteins was normalized to actin protein. The total load of the protein was 60 µg/µL. The dilution ratio of primary antibody was 1:1000, and that of secondary antibody was 1:5000. The type of secondary antibody was Goat anti-Rabbit. Chemiluminescence signals were quantified using a chemiluminescence imaging system (BIO RAD, Hong Kong, China).

Here, 1 × 10⁶ HepG2 cells were cultured in a 12-well plate, fixed in 4% paraformaldehyde for 30 min, and then blocked in PBS + 1% BSA for 1 h at room temperature. Following that, the cells were incubated with primary antibodies (ab61182, Abcam, Shanghai, China) at 4 °C overnight. On the next day, the cells were incubated with secondary fluorescent-conjugated IgG (FITC-IgG) (AS011, ABclonal, Woburn, MA, USA) at room temperature. The primary antibody dilution concentration was 1:200 and the secondary antibody dilution concentration was 1:200. After 1 h, the cells were washed with PBS for three times. DAPI was used to counterstain the nuclei. An inverted fluorescence microscope was employed to capture images. All experiments were repeated three times.