Hoechst 33,342/PI double staining confirmed that the proportion of PI-positive HepG2 cells transfected with pcDNA3.1-ATG2B used to be greater than that in the control group, and this proportion was once down-regulated after 3-MA remedy (Fig. 3C). In vitro, the ranges of the autophagy markers LC3 and NLRP3 have been proven in the liver of NASH cells, whilst 3-MA reversed the impact of pcDNA3.1-ATG2B (Fig. 3D). This evidence concerns the gene ATG2B and metabolic dysfunction-associated steatohepatitis.