The Hep3B2.1–7 (Hep3B) cell line was obtained from ATCC (HB-8064; ATCC, Manassas, VA); the HuH-7 cell line was from the laboratory of Dr. Greg Gores (Mayo Clinic, Rochester, MN). The Rab10 knockout fibroblasts were supplied by the laboratory of David James (University of Sydney, Australia). Cells were maintained in minimum essential medium (MEM) media containing 10% fetal bovine serum (FBS), 100 U/mL of penicillin, and 100 μg/mL of streptomycin (Life Technologies, Carlsbad, CA), and grown at 37°C in 5% CO2.
The rabbit Rab10 antibody was from Sigma-Aldrich (R8906; St. Louis, MO). The Rab10(4E2) antibody was from GeneTex (GTX82800; Irvine, CA). Dyn2 antibodies were prepared as detailed.(9 (link)) Alexa Fluor secondary antibodies were used for immunofluorescence along with ProLong Antifade reagent (Thermo Fisher Scientific, Waltham, MA). Horseradish peroxidase (HRP) secondary antibodies (Invitrogen) for western blotting analysis, detected using SuperSignal West Pico substrate (Thermo Fisher Scientific). Diacylglycerol O-acyltransferase (DGAT) inhibitors, were purchased from Sigma-Aldrich.
The rabbit Rab10 antibody was from Sigma-Aldrich (R8906; St. Louis, MO). The Rab10(4E2) antibody was from GeneTex (GTX82800; Irvine, CA). Dyn2 antibodies were prepared as detailed.(9 (link)) Alexa Fluor secondary antibodies were used for immunofluorescence along with ProLong Antifade reagent (Thermo Fisher Scientific, Waltham, MA). Horseradish peroxidase (HRP) secondary antibodies (Invitrogen) for western blotting analysis, detected using SuperSignal West Pico substrate (Thermo Fisher Scientific). Diacylglycerol O-acyltransferase (DGAT) inhibitors, were purchased from Sigma-Aldrich.
