Sds polyacrylamide gel electrophoresis page gel

The protein expression levels of LDLR in the HepG2 cells were determined by Western blot assay at Cosmo Genetech (Sungsoo, Seoul, Korea). Briefly, cell protein extracts were prepared using radioimmunoprecipitation assay buffer (Thermo Scientific, Pierce, Rockford, IL, USA). Twenty micrograms of protein lysate were separated by electrophoresis on a 10% sodium dodecyl-sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) gel (Bio-Rad, Hercules, CA, USA) and transferred to a nitrocellulose membrane (Hybond-C Extra; GE Healthcare, Piscataway, NJ, USA). Nonspecific binding was blocked with 5% milk and incubated with 1:1000 primary antibody (LDLR Rabbit-mAb, #ab52818, Abcam) overnight at 4 °C. The secondary antibodies (horseradish peroxidase [HRP]-conjugated mouse anti-rabbit IgG, #SC-2357, Santa Cruz) were diluted 1:3000 in a fresh blocking solution. Protein bands were detected using the SuperSignal West Femto Kit (Thermo Fisher Scientific Inc., Waltham, MA, USA).

Worms of each genotype (80–100 individuals of L2 stage worms) were collected and washed with M9 buffer and boiled in sodium dodecyl sulphate (SDS) sample buffer for 10 min and loaded onto SDS-polyacrylamide gel electrophoresis (PAGE) gel (Bio-Rad). A 1:500 dilution of rabbit anti-phospho-p38 MAPK (Cell Signaling, #9211) and a 1:10,000 dilution of mouse anti-actin (MPbio, #08691001) were used as primary antibodies. ImageJ was used to quantify the intensity of the immunoblot bands.