5000 Hela and HEK293T cells were seeded on day one in each well of the 96 well plate. Transfection of the constructs (M12-M22) with four different conditions were carried out on day two. The ratio of transfection is 1 (M construct) : 5 (stuffer/ miRsAg/ pcDNA5/ shRNA3) with a total amount of 50ng DNA.
Condition a: cotransfection with stuffer (salmon sperm DNA)
Condition b: cotransfection with synthetic RNA miRsAg
Condition c: cotransfection with empty pcDNA5
Condition d: cotransfection with synthetic shRNA3
A control consisting of the empty miMeasure plasmid (without binding site) was also cotransfected with the same conditions a, b, c and d.
The cells were used for measurements on day three.
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