Cyclophilin b

HepG2 human hepatoma cells, which represent the most used in vitro model to study liver metabolism and related disorders, were transiently transfected for 48 h by pooling 3 different target-specific siRNA oligo duplexes (MyBioSource, Inc., San Diego, CA, USA) of human HMBS gene (siPBGD) at a final concentration of 10 µM. Cyclophilin B (10 µM) was used as a scramble negative control (Horizon Discovery, Waterbeach, UK). To mimic fasting, both scramble and siPBGD cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM, Life Technologies-ThermoFisher Scientific, Waltham, MA, USA) with low glucose concentration (1.125 mg/L) for 6 h. A cultured medium of siPBGD cells was supplemented with 0.5 mM α-LA alone (siPBGD + α-LA) or in combination with 0.33 mM glucose (α-LA + Gluc) for 24 h before inducing fasting. Treatments were freshly prepared and administered when appropriate. The potential efficacy of α-LA alone and α-LA + Gluc were compared to 0.33 mM glucose (siPBGD + Gluc) [16 (link)].

Hep3B cells were transiently transfected for 48 h by pooling three different target-specific siRNA oligo duplexes directed against the exons 2, 4, and 5 of the human PNPLA3 (siHep3B) in order to improve the gene-silencing efficiency and at a final concentration of 10 μM (MyBioSource, Inc., San Diego, CA, USA). Cyclophilin B (10 μM) was used as a scramble negative control (Horizon Discovery, Waterbeach, UK).