Figure 2.

SIRT6 is the target of miR-125b and is up-regulated in HCCs. A. Dual-luciferase reporter assays. Sequences encoding wild-type and mutated fragments, mutated sequence, shown in the right panel, of the SIRT6 3’-UTR were inserted into a luciferase reporter plasmid. MiR-125b mimic was co-transfected with reporter plasmid into 293T cells. Relative renilla luciferase expression was normalized to firefly luciferase. B. Western blots shows the SIRT6 protein level in cells transfected with miR-125b mimic or control mimic. C. SIRT6 protein in tissue samples were measured by western blotting (adjacent non-tumor tissue (left lane) versus tumor tissue (right lane)). β-actin was used as loading control. D. Immunohistochemistry (IHC) shows the SIRT6 expression in adjacent non-tumor tissues (left) and tumor tissues (right). E. The level of SIRT6 mRNA in tissue samples was detected by qRT-PCR. Data were analyzed using 2^-ΔCT method in tissue samples and normalized to the expression of β-actin. F. SIRT6 expression in HCC patients and HTSeq-Counts data acquired from TCGA database, which is shown with a logarithmic conversion (log2^counts). Unpaired t-test. G. SIRT6 gene is related with the overall survival rate in HCC patients. Red line: high expression, blue line: low expression. Data is acquired from TCGA database and analyzed via Linked Omics bioinformatics [23].