Dual light luciferase and β gal reporter gene assay system

Cells were used lipofectamine 2000 (Thermo Scientific, Carlsbad, CA, USA) to cotransfected with luciferase reporter plasmids driven by Cx43 promoters (0.66 μg) 1 (link) and pTCYLacZ (0.34 μg), which is a β-galactosidase (β-gal) expression plasmid driven by the β-actin promoter. Then, cells were treated with EPA (0-100 μM) and cell lysates were harvested at different concentrations after 6 h transfection. The cell lysates were assessed for their luciferase activities as determined by a dual-light luciferase and β-gal reporter gene assay system (Promega, Madison, WI, USA) using a luminometer (Minilumate LB9506, Bad Wildbad) 3 (link). Relative luciferase activity was measured as luciferase activity divided by β-gal activity to normalize transfection efficiency per microgram protein. Meanwhile, the BCA protein assay (Pierce) was used to determine protein content in each sample.

Cells grown in 24-well plates were cotransfected with luciferase reporter plasmids driven by Cx43 promoters (0.66 μg) and pTCYLacZ (0.34 μg), a β-galactosidase (β-gal) expression plasmid driven by the β-actin promoter, by lipofectamine 2000 (Invitrogen). At 6 h post-transfection, cells were treated with resveratrol or S.C. and cell lysates were harvested at different concentrations. The cell lysates were assessed for their luciferase activities determined by a dual-light luciferase and β-gal reporter gene assay system (Promega, Madison, WI, USA) using a luminometer (Minilumate LB9506, Bad Wildbad). Relative luciferase activity was measured as luciferase activity divided by β-gal activity to normalize transfection efficiency per microgram protein. The protein content in each sample was determined by the BCA protein assay (Pierce).