Journal: Cancer Science

Article Title: NS 5 ABP 37 inhibits liver cancer by impeding lipogenesis and cholesterogenesis

doi: 10.1111/cas.13117

Figure Lengend Snippet: NS5ABP37 inhibits sterol regulatory element‐binding protein (SREBP) transcription activity in HepG2 and L02 hepatocellular carcinoma cells. HepG2 and L02 cells were transiently transfected with pENTER (normal control [NC]), pENTER‐NS5ABP37, siRNA control (siNC), or siRNA‐NS5ABP37. After 48 h, the mRNAand protein were extracted. The mRNA (A,B) and protein (C,D) levels of SREBP2, HMG‐CoA reductase (HMGCR), SREBP1c, and fatty acid synthase (FASN) were measured by RT‐PCR and Western blot analysis, respectively. Finally, HepG2 and L02 cells were transiently cotransfected with SREBP1c or SREBP2 promoter and plasmid pENTER, pENTER‐NS5ABP37, siRNA control, or siRNA‐NS5ABP37. After 24 h, SREBP1c and SREBP2 promoter activities were determined in a microplate luminometer (E,F). Data are mean ± SD from triplicate experiments. * P < 0.05, ** P < 0.005.

Article Snippet: SREBP1c or SREBP2 promoter activities were measured on a microplate luminometer, according to the technical manual for the Dual‐Luciferase Reporter Assay System (E1910; Promega).

Techniques: Binding Assay, Activity Assay, Transfection, Control, Reverse Transcription Polymerase Chain Reaction, Western Blot, Plasmid Preparation