Fig 4. In vivo luciferase assay study of HepG2 cells co-transfected with the reporter plasmids and different concentrations of miR-181a.

HepG2 cells were electroporated with 4ug of reporter plasmids and 10nM or 100nM miR-181a and seeded on 24 well plates. They were harvested after a 24h incubation and assayed for firefly and renilla luminescence using a manual luminometer. Results show that miR-181a binds to some extent, to the control vector, because of its dose dependent reduction in firefly luciferase activity upon transfection with the control vector. However, as compared to the control, miR-181a shows a significantly stronger binding to both the plasmids containing the 3’UTRs of CDKN1β and E2F7, with a further decrease in firefly luciferase activity by about two times when transfected with 10nM miR-181a and up to six times when transfected with 100nM miR-181a, as compared to when transfected with the control reporter plasmid.